HEMApSTR version 2.2

_{(version 2.2)}

Welcome to HEMApSTR!

Homepage of the Haemophilia A extragenic and intragenic factor VIII (F8) Short Tandem Repeats Map Inventory Database: A one-stop WWW resource site intended to provide free, easy access, in any location, to all researchers working primarily in Haemophilia A indirect carrier detection by PCR and linkage analysis on human distalmost Xq28 chromosomal band and, secondarily, in clinical and/or (new!) Forensic Applications of ChrX-STRs.

Keywords: Short Tandem Repeat; STRs; X-STRs; Xq28-STRs; ChrX-STRs; X-chromosome microsatellite; Simple Sequence Repeats-SSRs; polymorphic DNA marker.

The HEMApSTR URL is http://www.uenf.br/Uenf/Pages/CBB/LBT/HEMApSTR.html; hosted by the Laboratory of Biotechnology, Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Brazil.

Created by:

Prof. Dr. Enrique Medina-Acosta, M.Sc., Ph.D.

MyCVLattes - CNPq (Portuguese)
MyResearcherID (ISI Web of Knowledge)
MyScopusID
MyORCID
MyAcademia.edu

MyResearchGate

Filipe Brum Machado CVLattes, M.Sc., D.Sc. - CNPq (Portuguese).

Maintenance

Citing this resource

Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.

Contact information

Copyright and credits

Version updates (changes, clarifications, and WARNINGS!)

Table S1. Annotated genes present in the physical interval.

Table S2. Details about the In-Silico validation for the complete set of STR loci.

Table S3. Primers for amplification of the complete set of STR loci.

ATTENTION USERS!

HEMApSTR physical positions refer to NCBI36/hg18, build 36 version 3, NC_000023 human X-chromosome reference assembly coordinates (accession version NC_00023.9 gi:89161218). Therefore, caution must be exercised at mapping STR loci to within sequences from other builds (i.e., GRChH37/hg19). The user can utilize the Batch Coordinate Conversion (LiftOver) utility at UCSC Genome Bioinformatics Browser to convert genome coordinates and genome annotation files between assemblies. The current version supports both forward and reverse conversions, as well as conversions between selected species. Note that the Genomic Coordinate Position format is “chrX:start-end” (example chrX:34604027-34604065).

Table S1. Annotated genes present in the physical interval.

Annotated genes comprised in the physical interval spanning 2.394.042 bp on distalmost Xq28 band region (physical position 152121071 to 154515113 bp for the NCBI36/hg18, build 36 version 3, NC_000023 human X-chromosome reference sequence, accession version NC_00023.9 gi:89161218). A comprehensive table list of all 1324 protein CDS genes present in the updated human X-chromosome primary reference assembly sequence GRChH37/hg19 (gi:224589822) is available at NCBI protein CDS genes on X. The F8 gene is highlighted in red.

^(a)

NCBI Map Viewer locus name on human X-chromosome reference sequence with links that lead to gene information.

^(b)

Lengthand distance in base pairs from Xp-tel for the reference sequence NCBI36/hg18, build 36 version 3, NC_000023 human X-chromosome (accession version NC_00023.9 gi:89161218). Shown is the physical length spanned by each gene. We specifically referred to build 36.3 because the interpolation tool (Rutgers Map Interpolator) of the Rutgers second-generation combined linkage-physical map of the human genome currently uses NCBI36/hg18 coordinates ^[1].

^[1] Matise TC, Chen F, Chen W, De La Vega FM, Hansen M, He C et al. A second-generation combined linkage physical map of the human genome. Genome Res 2007; 17(12): 1783-86.

^(c)

Genes transcribed in the plus orientation (from the top down) or in the minus orientation (from bottom up).

^(d)

Online Mendelian Inheritance in Man accession number leads to a detailed description of the gene or disease.

^(e⁾

LIFEdb Xq28 accession leads to a systematic expression analysis of genes located on chromosomal arm Xq28 by Kolb-Kokocbinski et al. 2006 ^[2].

^[2] (Kolb-Kokocinski A, Mehrle A, Bechtel S, Simpson JC, Kioschis P, Wiemann S et al. The systematic functional characterisation of Xq28 genes prioritises candidate disease genes. BMC Genomics 2006; 7:29).

Table S1. Annotated genes present in the physical interval.

n^o		Gene symbol ^(a)	Physical interval ^(b)	Coding strand ^(c)	OMIM ^(d)	LIFEdb Xq28 ^(e)
1		MAGEA1	152134716-152139310	Minus	*300016	Available
2		LOC728317	152190272-152196454	Plus		Unavailable
3		LOC649201	152237415-152253805	Minus		Unavailable
4		ZNF275	152262060-152270249	Plus		Available
5		LOC649238	152315381-152317501	Minus		Unavailable
6		ZFP92	152329775-152340280	Plus		Available
7		TREX2	152363372-152365139	Minus	*300370	Available
8		UCHL5IP	152366318-152389283	Minus	*300540	Unavailable
9		LOC389904	152392294-152405740	Plus		Unavailable
10		BGN	152413605-152428198	Plus	*301870	Available
11		ATP2B3	152454774-152501581	Plus	*300014	Available
12		FAM58A	152506577-152517775	Minus		Available
13		DUSP9	152561182-152569971	Plus	*300134	Available
14		PNCK	152588417-152592974	Minus	*300680	Available
15		SLC6A8	152606586-152615234	Plus	#300352 / *300036	Available
16		BCAP31	152619146-152643081	Minus	*300398	Available
17		ABCD1	152643530-152663375	Plus	*300371 / #300100	Available
18		PLXNB3	152682905-152697989	Plus	*300214	Available
19		SRPK3	152699704-152704381	Plus		Available
20		IDH3G	152704415-152713161	Minus	*300089	Available
21		SSR4	152713288-152717148	Plus	*300090	Available
22		PDZD4	152720817-152749197	Minus	*300634	Available
23		L1CAM	152780581-152794505	Minus	*308840 / #307000 / #304100 / #303350 / #303350	Available
24		LCAP	152799321-152807638	Plus		Unavailable
25		AVPR2	152823564-152825834	Plus	#304800 / #300539 / *300538	Available
26		ARHGAP4	152826025-152844892	Minus	*300023	Available
27		ARD1A	152848571-152853662	Minus	*300013	Available
28		RENBP	152853917-152863426	Minus	*312420	Available
29		HCFC1	152866202-152890013	Minus	*300019	Available
30		TMEM187	152891185-152901840	Plus	*300059	Available
31		IRAK1	152929151-152938536	Minus	*300283	Available
32		MECP2	152940458-153016323	Minus	#312750 / #300673 / #300496 / #300260 / #300055 / *300005 / #105830	Available
33		LOC100128952	153013619-153016356	Plus		Unavailable
34		OPN1LW	153062939-153077701	Plus	+303900 / %303700	Available
35		OPN1MW	153101361-153114725	Plus	+303800 / %303700	Available
36		OPN1MW2	153138461-153151953	Plus		Unavailable
37		TEX28	153152126-153176632	Minus	*300092	Available
38		TKTL1	153177345-153211894	Plus	*300044	Available
39		FLNA	153230091-153252845	Minus	#311300 / #309350 / #304120 / #300537 / #300049 / +300017	Available
40		LOC100131857	153254916-153255347	Minus		Unavailable
41		EMD	153260981-153263075	Plus	#310300 / *300384	Available
42		RPL10	153279912-153283874	Plus	*312173	Available
43		SNORA70	153281816-153281950	Plus		Unavailable
44		DNASE1L1	153282773-153293621	Minus	*300081	Available
45		TAZ	153293071-153303259	Plus	#302060 / *300394 / #300183 / #300069	Available
46		ATP6AP1	153310172-153318056	Plus	*300197	Available
47		GDI1	153318715-153325008	Plus	%309541 / +300104	Available
48		FAM50A	153325702-153332190	Plus	*300453	Available
49		PLXNA3	153339817-153355179	Plus	*300022	Available
50		LAGE3	153358435-153360790	Minus	*300060	Available
51		UBL4A	153365250-153368126	Minus	*312070	Available
52		SLC10A3	153368842-153372189	Minus	*312090	Available
53		FAM3A	153387700-153397567	Minus	*300492	Available
54		G6PD	153412800-153428981	Minus	+305900	Available
55		IKBKG	153423653-153446455	Plus	#308300 / #300636 / #300584 / #300301 / #300291 / *300248	Available
56		LOC643894	153449493-153463455	Plus		Unavailable
57		LOC100132967	153452673-153453385	Plus		Unavailable
58		CTAG1A	153466610-153468269	Plus	*300657	Unavailable
59		CTAG1B	153499059-153500714	Minus	*300156	Available
60		CXorf52	153513930-153514642	Minus		Available
61		CTAG2	153533445-153535036	Minus	*300396	Available
62		GAB3	153556720-153632542	Minus	*300482	Available
63		DKC1	153644344-153659154	Plus	#305000 / #300240 / *300126	Available
64		SNORA36A	153649997-153650128	Plus		Available
65		SNORA56	153656467-153656595	Plus		Unavailable
66		MPP1	153660162-153686957	Minus	*305360	Available
67		LOC100132963	153704817-153717173	Minus		Unavailable
68		F8	153717260-153904192	Minus	+306700	Available
69		H2AFB1	153766511-153767027	Plus		Unavailable
70		F8A1	153767829-153769530	Plus	*305423	Available
71		FUNDC2	153908258-153938385	Plus		Available
72		MTCP1	153943080-153952830	Minus	*300116	Available
73		BRCC3	153952904-154004543	Plus	*300617	Available
74		VBP1	154097744-154121292	Plus	*300133	Available
75		RAB39B	154140720-154147046	Minus		Available
76		CLIC2	154158694-154217180	Minus	*300138	Available
77		H2AFB2	154263630-154263977	Plus		Unavailable
78		F8A2	154264958-154266073	Plus		Unavailable
79		F8A3	154340340-154341455	Minus		Unavailable
80		H2AFB3	154342274-154342790	Minus	*300445	Available
81		TMLHE	154372967-154495791	Minus		Available


Table S2. Details about the In-Silico validation for the complete set of STR loci included in comprehensive STR physical map inventory: *characteristic features of the F8 intragenic and extragenic STR loci* validated In-Silico and interpolated genetic distances**.
^(a)	Locus name: Newly described loci are named by the four letters acronym HEMA (Haemophilia A), followed by a number that indicates the distance in base pairs from Xp-tel. For example, HEMA152128.2 is a STR element and candidate locus located at 152.128.251 bp on NCBI36/hg18, build 36 version 3, NC_000023 human X-chromosome reference sequence, accession version NC_00023.9 gi:89161218. When the STR element is intragenic, we designated it to the intron (Int) or exon (Exon) number of the gene. When more than one STR element is found within a given intron or exon, a dot one digit is included to indicate the order of the elements in the intron or the exon (for example, TMLHEInt1.3 refers to the third (“.3”) STR element located within intron 1 (“Int1”) of the TMLHE gene). Known loci are named as published, when genotyped (i.e., tried-and-tested), and/or by their designated Sequence Tagged Site (UniSTS) code. A UniSTS entry is a short DNA segment that occurs only once in the human genome and whose exact location and order of bases are known. Because (in principle) each short DNA segment listed at UniSTS is unique, STSs are helpful for chromosome placement of mapping and sequencing data from many different laboratories. STSs serve as landmarks on the physical map of the human genome (UniSTS is a National Center for Biotechnology Information NCBI resource). The loci listed are hyperlinked to allow navigation to the reference locus sequences and the map coordinates of suitable ampliprimer sequences.
^(b)	Core, consensus sequence pattern of the repeat element (upper strand).
^(c)	Distance in base pairs from Xp-tel, as determined using the reference sequence NCBI36/hg18, build 36 version 3, NC_000023 human X-chromosome reference sequence, accession version NC_00023.9 gi:89161218, assembly of human X-chromosome. Shown is the interval distance for each STR element. The F8 gene maps on the interval 153717260-153904192 bp.
^(d)	Repeat copy number.
^(e)	Percentage of identity between repeat copies (Note: the stand-alone and the online versions of the Tandem Repeats Finder program report “percent match between adjacent copies”; the online Tandem Repeats Database reports “percent match between a consensus sequence and each copy”).
^(f)	Minimum alignment score value to list the selected STR loci depended on the size of the repeat element size. For dinucleotide STR elements, the minimum score was 54, which corresponds to the lowest score for any of the F8 intragenic STR known to be polymorphic by direct genotyping; that is, the REN91235(F8Int6) locus. For trinucleotide, tetranucleotide and pentanucleotide X-STRs elements, the minimum score values shown to be polymorphic by In-Silico validation were 80, 66, and 52, respectively. For the F8 gene (region highlighted in red), however, all dinucleotide and trinucleotide X-STRs elements with score > 30 are listed. All loci listed present minimum percent matches > 80. STR elements located in pseudogenes and copy number variations (CNVs, structural large-scale variation in the human genome) were excluded from the inventory.
^(g)	In-Silico validated polymorphism index (IPI, or genetic variation) obtained by comparison of three reference X-chromosome sequences, available at NCBI with the accession numbers NC_000023 (reference assembly), AC_000066 (alternate assembly, based on Celera assembly), AC_000155 (alternate assembly, based on HuRef). (Note: At least six other individual genome sequences (haploid and diploid) are currently available from public databases (Asian, Southern African Bushman (Complete Khoisan and Bantu genomes from southern Africa and James Watson’s Genome); search date through February 15, 2010) and promising prospects of a deep catalogue of human genetic variation comprising 1000 individual genomes; we will make efforts to update the polymorphism indexes, as relevant STR information becomes available). For instance, validation indexes that include information about allelic variants sequenced in the southern African Bushman^[1] are highlighted in green (i.e., 3); Validation indexes that include new allelic variants sequenced in the southern African Bushman are highlighted in cyan (i.e., 1). ^[1]Schuster SC, Miller W, Ratan A, Tomsho LP, Giardine B, Kasson LR, et al. Complete Khoisan and Bantu genomes from southern Africa. Nature; 463(7283): 943-7.
^(h)	Interpolatedgenetic distances in centiMorgans, determined using the online regression-based smoothed Rutgers Map v.2 interpolator of the human genome. The Rutgers map includes the largest set of polymorphic markers for which genotype data are publicly available that combines genotype data from both the CEPH and deCODE pedigrees (for some markers), that incorporates SNPs, and that incorporates sequence-based positional information. The interpolated genetic distance in females between the interval border markers DXS9897 and DXS1108 is 3.22 cM (i.e. 3.22% expected rate of recombination). The interpolated genetic distance from DXS9897 to F8Int1 (the upstream most STR element within the F8 gene) is 14.33 times the genetic distance from REN90932 (F8Int25.3; the downstream most STR element within the F8 gene) and DXS1108. The risk of recombination 3’ to the F8 gene is thus 3.27 times the risk of recombination 5’ to the F8 gene for STR markers equidistant 783.522 bp of the F8 gene.
⁽ⁱ⁾	Validated In-Silico only in two reference sequences of X-chromosome, since no sequence information for this STR element was available in all three reference sequences.
^(j)	The REN nomenclature to define UniSTS comes from a set of primer pairs that was developed in Dr. Bing Ren lab for use in the ENCyclopedia of DNA Elements (ENCODE) project^[2]. ^[2]The ENCODE (ENCyclopedia Of DNA Elements) Project. Science 2004; 306(5696): 636-40.

Table S2. Details about the In-Silico validation for the complete set of STR loci.

n^o	*Locus* name ^(a)	Consensus Pattern ^(b)	Physical Position ^(c)	Copy Number ^(d)	Percent Matches ^(e)	Match Score ^(f)	Validation index ^(g)	Map length (cM) ^(h)
1	DXS9897 New!	CTAT	152121071--152121128	14.8	96	109	2	192.76
2	HEMA152128.2	GT	152128251--152128283	16.5	100	66	2	192.78
3	HEMA152141.6	AG	152141683--152141709	13.5	100	54	3	192.81
4	HEMA152149.9	TATT	152149956--152149998	11.0	95	79	2	192.84
5	HEMA152155.2	AC	152155298--152155326	14.5	100	58	2	192.86
6	HEMA152155.6	TG	152155655--152155695	20.5	89	64	2	192.86
7	LOC649201Int2	GT	152250679--152250715	18.5	100	74	3	193.16
8	HEMA152257.3	TG	152257313--152257369	28.5	100	114	2	193.17
9	DXS15	CA	152285381--152285422	21.0	100	84	1⁽ⁱ⁾	193.25
10	HEMA152317.9	TG	152317955--152317988	17.5	93	61	1	193.33
11	HEMA152342.0	AC	152342046--152342085	20.0	100	80	2	193.39
12	DXS1356	AGGG	152351238--152351343	11.3	90	72	1⁽ⁱ⁾	193.42
12	DXS1356	GGAA	152351238--152351343	16.0	100	128	1⁽ⁱ⁾	193.42
13	UCHL5IPInt8	GGGA	152373164--152373200	9.3	100	74	1⁽ⁱ⁾	193.48
14	DXS9901(BGNExon8)	CT	152427681--152427758	15.0	100	60	2	193.63
14	DXS9901(BGNExon8)	TG	152427681--152427758	22.5	100	90	2	193.63
15	HEMA152442.8	GT	152442878--152442911	17.0	93	59	1	193.67
16	ATP2B3Int20	AC	152489656--152489691	18.0	100	72	2	193.80
17	HEMA152503.2	AAGG	152503201--152503236	9.0	100	72	2	193.84
18	HEMA152520.9	TTAT	152520968--152521000	8.3	100	66	1	193.89
19	HEMA152524.1	AAGA	152524121--152524353	56.5	88	285	3	193.90
20	HEMA152539.8	AGAAA	152539891--152539990	20.2	89	112	1⁽ⁱ⁾	193.93
21	DXS8087(SHGC-152185)	TG	152547714--152547748	17.5	100	70	2	193.94
22	HEMA152581.8 *WARNING!*	AAAAT	152581889--152581940	10.6	93	79	1	194.01
23	PNCKInt2	AC	152591604--152591641	18.0	89	58	1	194.02
24	SLC6A8Exon1	GCC	152606681--152606964	11.0	93	57	1⁽ⁱ⁾	194.05
24	SLC6A8Exon1	GCC	152606681--152606964	16.0	77	51	1⁽ⁱ⁾	194.05
25	PDZD4Int1	TC	152741273--152741325	26.5	84	70	2	194.32
26	REN87871^(j)*WARNING!*	TTTC	152769114--152769164	12.8	100	102	2	194.40
27	HEMA152778.1	AAGG	152778106--152778183	19.0	92	129	1	194.42
28	REN87914	TTTC	152779117--152779411	45.3	83	185	3	194.42
28	REN87914	CTCT	152779117--152779411	24.0	91	102	3	194.42
29	DXS9796(REN88038)	GT	152807881--152807923	12.5	91	41	1	194.50
29	DXS9796(REN88038)	GA	152807881--152807923	9.5	100	38	1	194.50
30	REN88052	TC	152810851--152810880	15.0	100	60	2	194.50
31	REN88058	AAAG	152812156--152812299	36.8	82	176	2	194.51
32	RENBPInt9.1	AAAT	152855845--152855891	11.8	100	94	1	194.61
33	REN88388(TMEM187Int1.1)	AC	152892330--152892356	13.5	100	54	2	194.70
34	TMEM187Int1.2	TTTTG	152897398--152897446	9.8	100	98	2⁽ⁱ⁾	194.71
35	REN88442	TATTT	152910286--152910340	11.2	96	103	1	194.74
36	HEMA152913.2	GT	152913227--152913428	101.0	80	224	2	194.74
37	REN88627(MECP2Int2.3)	AT	152955789--152955831	21.5	100	86	2	194.83
38	REN88666/HSC270_(M3)/ (MECP2Int2.2)	GT	152964545--152964591	23.5	100	94	3	194.85
39	REN88881	AG	153019029--153019057	14.5	100	58	2	194.96
40	HEMA153020.0	GATA	153020002--153020061	15.0	92	66	2	194.96
41	REN88942	GT	153034597--153034664	34.5	86	93	2	194.99
42	REN88956	TG	153038190--153038232	21.5	100	86	1	195.00
43	HEMA153047.7	AC	153047747--153047824	15.5	100	62	3	195.02
43	HEMA153047.7	AT	153047747--153047824	24.0	95	87	3	195.02
44	HEMA153050.0	TG	153050087--153050120	17.0	100	68	1	195.02
45	TKTL1Int1	TTTA	153184723--153184773	12.8	100	102	2	195.29
46	REN89483(TAZInt5)	ATAG	153296967--153297133	41.0	82	95	1	195.47
47	HEMA153333.1	TTCC	153333110--153333151	10.8	89	68	1	195.52
48	G65800	TGTT	153376711--153376758	12.3	91	80	1	195.56
49	REN89 870(FAM3AInt2)	AAG	153393770--153393811	14.0	100	84	2	195.58
50	REN89898/REN89899 New! *WARNING!*	AT	153401350--153401379	15.0	100	60	1	195.59
51	stSG604115/REN89922	TG	153408151--153408189	19.5	89	60	1	195.60
52	REN90010/GDB:179459/ (IKBKGInt1) New!	TG	153431644--153431687	22.0	95	79	1	195.62
53	stSG604170/REN90051/ REN90052	GA	153477086--153477279	103.0	81	225	1	195.65
54	DXS1073(REN90074) New! *WARNING!*	TG	153482143--153482182	20.0	100	80	2	195.65
55	REN90085	TTAT	153484584--153484617	8.5	100	68	1	195.65
56	REN90200(HA472) New!	CTT	153553700--153553902	67.0	94	343	3	195.69
57	stSG604240/REN90371 (GAB3Int2.2) New! WARNING!	AT	153595212--153595255	22.0	100	88	2	195.70
58	stSG604242/REN90378 (GAB3Int2.1)	TG	153596862--153596924	17.0	81	41	1	195.70
58	stSG604242/REN90378 (GAB3Int2.1)	AG	153596862--153596924	13.5	100	54	1	195.70
59	stSG604248/REN90414 (GAB3Int1) New!	TAAA	153605812--153605855	11.0	100	88	1	195.70
60	stSG604294/REN90682/ UniSTS:68640/HSC269_(M1) (MPP1Int5) *WARNING!*	AC	153668571--153668616	23.0	100	92	2⁽ⁱ⁾	195.74
61	REN90833 New! WARNING!	TTAT	153694353--153694393	10.3	100	82	2	195.75
	3’ end of the F8 gene		Ends at 153717260 (minus strand)					195.77
62	REN90932(F8Int25.3) New!	TG	153731553--153731589	18.5	100	74	1	195.77
63	stSG604314/REN90841 (F8Int25.2) New!	TG	153736060--153736088	14.5	100	58	2	195.77
64	REN90957(F8Int25.1) New!	AG	153740631--153740651	10.5	89	33	1	195.77
65	REN90965(F8Int24.1) New!	TCCT	153742466--153742535	12.3	82	44	1	195.77
65	REN90965(F8Int24.1) New!	TC	153742466--153742535	9.5	100	38	1	195.77
66	F8Int22 New!	GT	153757206--153757243	19.0	100	76	2	195.78
67	HSC271_(M4)/F8Int21 New!	AC	153777976--153778027	26.0	92	86	1	195.78
68	REN91100(F8Int13.2) New!	AC	153817451--153817493	21.5	100	86	2	195.80
69	REN91114(F8Int13.1)	AT	153820354--153820370	8.5	100	34	2	195.80
70	stSG604366/REN91179/ REN91180(F8Int10)	GA	153842062--153842082	10.5	100	42	1	195.81
71	stSG604368(F8Int9.2) New!	AG	153844339--153844390	25.5	92	86	1	195.81
72	stSG604369/REN91192 (F8Int9.1) New!	CA	153845551--153845578	14.0	92	47	1	195.81
73	REN91235(F8Int6) New!	TG	153855435--153855461	13.5	100	54	1	195.81
74	stSG604384/REN91318 (F8Int3)	TTG	153874731--153874753	7.7	100	46	1	195.82
75	F8Int1 New!	AC	153884082--153884115	17.0	100	68	2⁽ⁱ⁾	195.83
	5’ end of the F8 gene		Starts at 153904192 (minus strand)					195.84
76	HEMA154006.7	AC	154006702--154006794	46.5	91	96	2	195.87
77	stSG604469 New! *WARNING!*	AT	154019738--154019777	20.0	100	80	3	195.87
78	stSG604486 New! *WARNING!*	AC	154050935--154050987	26.5	100	106	2	195.88
79	VBP1Int4	GT	154116577--154116620	22.0	95	79	1	195.91
80	HEMA154126.0	TG	154126085--154126111	13.5	100	54	2	195.91
81	HEMA154130.5 New! *WARNING!*	CT	154130572--154130609	19.0	100	76	1	195.91
82	CLIC2Int1	AT	154208272--154208313	21.0	100	84	2	195.93
83	HEMA154229.0 *WARNING!*	TTTC	154229069--154229113	11.8	95	76	2	195.93
84	HEMA154279.5(HA544)	TTC	154279540--154279610	23.7	100	142	2⁽ⁱ⁾	195.94
85	HEMA154281.2	AC	154281204--154281236	16.5	93	57	1	195.94
86	HEMA154311.3 New!	GAAGA	154311336--154311361	5.2	100	52	2	195.95
87	TMLHEInt5 New!	TTTTA	154393088--154393119	6.4	100	64	2⁽ⁱ⁾	195.97
88	TMLHEInt2 New! WARNING!	GT	154415524--154415567	22.0	100	88	2⁽ⁱ⁾	195.97
89	TMLHEInt1.3 New!	ATTC	154436837--154436875	9.8	100	78	3	195.97
90	TMLHEInt1.2	AC	154468260--154468308	24.5	95	89	2	195.98
91	TMLHEInt1.1 New! WARNING!	GAAA	154486183--154486266	20.5	90	125	2⁽ⁱ⁾	195.98
92	HEMA154498.9 New!	TC	154498956--154499028	38.0	91	125	2	195.98
93	HEMA154507.3 New!	ATCT	154507351--154507428	20.5	87	119	3	195.98
94	DXS1108 New!	CA	154515075--154515113	19.5	94	69	2	195.98

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS9897	NC_000023	152121071--152121128	14.8	96	109
	AC_000066	152695987--152696044	14.8	96	109
	AC_000155	141127878--141127931	13.8	96	101

New!

This locus has recently been tried-and-tested as informative using the pair of primers here provided^[1].

^[1]

Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.

DXS9897	Information
Amplicon	232-236 bp
Forward ampliprimer	TTCTGCTGTGCAATACATCTGA
Reverse ampliprimer	CAGCAGACATTATTGAGGGAGA

>ref|NC_000023.9|

TTCTGCTGTGCAATACATCTGACTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCATCTATCTAGCCATCTACC

TACCTACCTATCATATATAGAAACACTATCTCTATGACTTTGATTCATTTAGCTGCTACTTCATCTGCTTCAATGATTTAAAAAGAACAT

CTTGACTTCATTTCTTGATCTCATCTTTCTGTTTTCTCCCTCAATAATGTCTGCTG

>ref|AC_000066.1|

TTCTGCTGTGCAATACATCTGACTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCATCTATCTAGCCATCTACC

TACCTACCTATCATATATAGAAACACTATCTCTATGACTTTGATTCATTTAGCTGCTACTTCATCTGCTTCAATGATTTAAAAAGAACAT

CTTGACTTCATTTCTTGATCTCATCTTTCTGTTTTCTCCCTCAATAATGTCTGCTG

>ref|AC_000155.1|

TTCTGCTGTGCAATACATCTGACTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCATCTATCTAGCCATCTACCTACC

TACCTATCATATATAGAAACACTATCTCTATGACTTTGATTCATTTAGCTGCTACTTCATCTGCTTCAATGATTTAAAAAGAACATCTTG

ACTTCATTTCTTGATCTCATCTTTCTGTTTTCTCCCTCAATAATGTCTGCTG

DXS9897

In-Silico polymorphism validation

Sequence 1: ref_NC_000023 236 bp

Sequence 2: ref_AC_000066 236 bp

Sequence 3: ref_AC_000155 232 bp

ref_NC_000023 TTCTGCTGTGCAATACATCTGACTATCTATCTATCTATCTATCTATCTATCTATCTATCT

ref_AC_000066 TTCTGCTGTGCAATACATCTGACTATCTATCTATCTATCTATCTATCTATCTATCTATCT

ref_AC_000155 TTCTGCTGTGCAATACATCTGACTATCTATCTATCTATCTATCTATCTATCTATCTATCT

************************************************************

ref_NC_000023 ATCTATCTATCATCTATCTAGCCATCTACCTACCTACCTATCATATATAGAAACACTATC

ref_AC_000066 ATCTATCTATCATCTATCTAGCCATCTACCTACCTACCTATCATATATAGAAACACTATC

ref_AC_000155 ATCTATC----ATCTATCTAGCCATCTACCTACCTACCTATCATATATAGAAACACTATC

******* *************************************************

ref_NC_000023 TCTATGACTTTGATTCATTTAGCTGCTACTTCATCTGCTTCAATGATTTAAAAAGAACAT

ref_AC_000066 TCTATGACTTTGATTCATTTAGCTGCTACTTCATCTGCTTCAATGATTTAAAAAGAACAT

ref_AC_000155 TCTATGACTTTGATTCATTTAGCTGCTACTTCATCTGCTTCAATGATTTAAAAAGAACAT

************************************************************

ref_NC_000023 CTTGACTTCATTTCTTGATCTCATCTTTCTGTTTTCTCCCTCAATAATGTCTGCTG

ref_AC_000066 CTTGACTTCATTTCTTGATCTCATCTTTCTGTTTTCTCCCTCAATAATGTCTGCTG

ref_AC_000155 CTTGACTTCATTTCTTGATCTCATCTTTCTGTTTTCTCCCTCAATAATGTCTGCTG

********************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152128.2	NC_000023	152128251--152128283	16.5	100	66
	AC_000066	152688832--152688864	16.5	100	66
	AC_000155	141134872--141134906	17.5	100	70

HEMA152128.2	Information
Amplicon	220-222 bp
Forward ampliprimer	GCACATACAGCCTTCAAGAGG
Reverse ampliprimer	CGATGAAAAGTCACTGCGTCT

>ref|NC_000023.9|

GCACATACAGCCTTCAAGAGGACTTTGCTCCAGCTCTGCAAAGTATTATCAGCTAGCTGGGGCAGTCACTGTGACTTCAAAATGTCATTT

CACTGTTATTTTATTATGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGACGTTAAGCAAAGATCTTTGTTTTCTTTCCTCTCATAT

TTCCTTATCAGGTAACGTAAGACGCAGTGACTTTTCATCG

>ref|AC_000066.1|

GCACATACAGCCTTCAAGAGGACTTTGCTCCAGCTCTGCAAAGTATTATCAGCTAGCTGGGGCAGTCACTGTGACTTCAAAATGTCATTT

CACTGTTATTTTATTATGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGACGTTAAGCAAAGATCTTTGTTTTCTTTCCTCTCATAT

TTCCTTATCAGGTAACGTAAGACGCAGTGACTTTTCATCG

>ref|AC_000155.1|

GCACATACAGCCTTCAAGAGGACTTTGCTCCAGCTCTGCAAAGTATTATCAGCTAGCTGGGGCAGTCACTGTGACTTCAAAATGTCATTT

CACTGTTATTTTATTATGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGACGTTAAGCAAAGATCTTTGTTTTCTTTCCTCTCAT

ATTTCCTTATCAGGTAACGTAAGACGCAGTGACTTTTCATCG

HEMA152128.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 220 bp

Sequence 2: ref|AC_000066.1| 220 bp

Sequence 3: ref|AC_000155.1| 222 bp

ref|NC_000023.9| GCACATACAGCCTTCAAGAGGACTTTGCTCCAGCTCTGCAAAGTATTATCAGCTAGCTGG

ref|AC_000066.1| GCACATACAGCCTTCAAGAGGACTTTGCTCCAGCTCTGCAAAGTATTATCAGCTAGCTGG

ref|AC_000155.1| GCACATACAGCCTTCAAGAGGACTTTGCTCCAGCTCTGCAAAGTATTATCAGCTAGCTGG

************************************************************

ref|NC_000023.9| GGCAGTCACTGTGACTTCAAAATGTCATTTCACTGTTATTTTATTATGAGTGTGTGTGTG

ref|AC_000066.1| GGCAGTCACTGTGACTTCAAAATGTCATTTCACTGTTATTTTATTATGAGTGTGTGTGTG

ref|AC_000155.1| GGCAGTCACTGTGACTTCAAAATGTCATTTCACTGTTATTTTATTATGAGTGTGTGTGTG

************************************************************

ref|NC_000023.9| TGTGTGTGTGTGTGTGTGTGTG--ACGTTAAGCAAAGATCTTTGTTTTCTTTCCTCTCAT

ref|AC_000066.1| TGTGTGTGTGTGTGTGTGTGTG--ACGTTAAGCAAAGATCTTTGTTTTCTTTCCTCTCAT

ref|AC_000155.1| TGTGTGTGTGTGTGTGTGTGTGTGACGTTAAGCAAAGATCTTTGTTTTCTTTCCTCTCAT

********************** ************************************

ref|NC_000023.9| ATTTCCTTATCAGGTAACGTAAGACGCAGTGACTTTTCATCG

ref|AC_000066.1| ATTTCCTTATCAGGTAACGTAAGACGCAGTGACTTTTCATCG

ref|AC_000155.1| ATTTCCTTATCAGGTAACGTAAGACGCAGTGACTTTTCATCG

******************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152141.6	NC_000023	152141683--152141709	13.5	100	54
	AC_000066	152675407--152675431	12.5	100	50
	AC_000155	141148183--141148211	14.5	100	58

HEMA152141.6	Information
Amplicon	259-261 bp
Forward ampliprimer	GGCAGCCACAGGTAAGACAT
Reverse ampliprimer	CTTGAACTTGGGAGGTGGAG

>ref|NC_000023.9|

GGCAGCCACAGGTAAGACATAATTTGTACATAATTGTACATAATTGAAGCACATTTGACATTTTTACAGTGTTCTCTTTCTCATCAATGA

ATATGCTATGCTCCTCTATAATATGTGTGTGTGAGTGTGTGTATATATATATATATATATAGAGAGAGAGAGAGAGAGAGAGAGAGAAAG

ATGGCATCTCCCTCTGTCTACCAGGCTGGAGTGCAGTGGCACTATCTCGGCTCACTGTAATCTCCACCTCCCAAGTTCAAG

>ref|AC_000066.1|

GGCAGCCACAGGTAAGACATAATTTGTACATAATTGTACATAATTGAAGCACATTTGACATTTTTACAGTGTTCTCTTTCTCATCAATGA

ATATGCTATGCTCCTCTATAATATGTGTGTGTGAGTGTGTGTTATATATATATATATATATAGAGAGAGAGAGAGAGAGAGAGAGAAAGA

TGGCATCTCCCTCTGTCTACCAGGCTGGAGTGCAGTGGCACTATCTCGGCTCACTGTAATCTCCACCTCCCAAGTTCAAG

>ref|AC_000155.1|

GGCAGCCACAGGTAAGACATAATTTGTACATAATTGTACATAATTGAAGCACATTTGACATTTTTACAGTGTTCTCTTTCTCATCAATGA

ATATGCTATGCTCCTCTATAATATGTGTGTGTGAGTGTGTGTATATATATATATATAGAGAGAGAGAGAGAGAGAGAGAGAGAGAAAGAT

GGCATCTCCCTCTGTCTACCAGGCTGGAGTGCAGTGGCACTATCTCGGCTCACTGTAATCTCCACCTCCCAAGTTCAAG

HEMA152141.6

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 261 bp

Sequence 2: ref|AC_000066.1| 260 bp

Sequence 3: ref|AC_000155.1| 259 bp

ref|AC_000066.1| GGCAGCCACAGGTAAGACATAATTTGTACATAATTGTACATAATTGAAGCACATTTGACA

ref|NC_000023.9| GGCAGCCACAGGTAAGACATAATTTGTACATAATTGTACATAATTGAAGCACATTTGACA

ref|AC_000155.1| GGCAGCCACAGGTAAGACATAATTTGTACATAATTGTACATAATTGAAGCACATTTGACA

************************************************************

ref|AC_000066.1| TTTTTACAGTGTTCTCTTTCTCATCAATGAATATGCTATGCTCCTCTATAATATGTGTGT

ref|NC_000023.9| TTTTTACAGTGTTCTCTTTCTCATCAATGAATATGCTATGCTCCTCTATAATATGTGTGT

ref|AC_000155.1| TTTTTACAGTGTTCTCTTTCTCATCAATGAATATGCTATGCTCCTCTATAATATGTGTGT

************************************************************

ref|AC_000066.1| GTGAGTGTGTGT-TATATATATATATATATATAGAGAGAGAGAGAGAGAGAGAGAGAAAG

ref|NC_000023.9| GTGAGTGTGTGTATATATATATATATATATAGAGAGAGAGAGAGAGAGAGAGAGAGAAAG

ref|AC_000155.1| GTGAGTGTGTGT--ATATATATATATATAGAGAGAGAGAGAGAGAGAGAGAGAGAGAAAG

************ *************** * ****************************

ref|AC_000066.1| ATGGCATCTCCCTCTGTCTACCAGGCTGGAGTGCAGTGGCACTATCTCGGCTCACTGTAA

ref|NC_000023.9| ATGGCATCTCCCTCTGTCTACCAGGCTGGAGTGCAGTGGCACTATCTCGGCTCACTGTAA

ref|AC_000155.1| ATGGCATCTCCCTCTGTCTACCAGGCTGGAGTGCAGTGGCACTATCTCGGCTCACTGTAA

************************************************************

ref|AC_000066.1| TCTCCACCTCCCAAGTTCAAG

ref|NC_000023.9| TCTCCACCTCCCAAGTTCAAG

ref|AC_000155.1| TCTCCACCTCCCAAGTTCAAG

*********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152149.9	NC_000023	152149956--152149998	11.0	95	79
	AC_000066	152667118--152667160	11.0	95	79
	AC_000155	141156458--141156500	11.0	95	79

HEMA152149.9	Information
Amplicon	260 bp
Forward ampliprimer	TGGGCTTTATTCCCTTCCAT
Reverse ampliprimer	TGACGAGTTAGTGGGTGCAG

>ref|NC_000023.9|

TGGGCTTTATTCCCTTCCATTGATATATTCTTATAATAATGATAATTCCGACTACACTGATCATTGTTAGTTTATAAGCCTTGAAATGAA

GTAGGGTAACCCCACCACATTTGTTCTATTTTTTATTTATTTATTTATTTATTTATTTATTTATTTATTATTATTATACTTTAAGTTTTA

GGGTACATGTGCACAATGTGCAGGTTAGTTACACATGTATACATGTGCCATGCTGGTGCGCTGCACCCACTAACTCGTCA

>ref|AC_000066.1|

TGGGCTTTATTCCCTTCCATTGATATATTCTTATAATAATGATAATTCCGACTACACTGATCATTGTTAGTTTATAAGCCTTGAAATGAA

GTAGGGTAACCCCACCACATTTGTTCTATTTTTTATTTATTTATTTATTTATTTATTTATTTATTTATTATTATTATACTTTAAGTTTTA

GGGTACATGTGCACAATGTGCAGGTTAGTTACACATGTATACATGTGCCATGCTGGTGCGCTGCACCCACTAACTCGTCA

>ref|AC_000155.1|

TGGGCTTTATTCCCTTCCATTGATATATTCTTATAATAATGATAATTCCGACTACACTGATCATTGTTAGTTTATAAGCCTTGAAATGAA

GTAGGGTAACCCCACCACATTTGTTCTATTTTTTATTTATTTATTTATTTATTTATTTATTTATTTATTATTATTATACTTTAAGTTTTA

GGGTACATGTGCACAATGTGCAGGTTAGTTACACATGTATACATGTGCCATGCTGGTGCGCTGCACCCACTAACTCGTCA

HEMA152149.9

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 260 bp

Sequence 2: ref|AC_000066.1| 260 bp

Sequence 3: ref|AC_000155.1| 260 bp

ref|AC_000066.1| TGGGCTTTATTCCCTTCCATTGATATATTCTTATAATAATGATAATTCCGACTACACTGA

ref|NC_000023.9| TGGGCTTTATTCCCTTCCATTGATATATTCTTATAATAATGATAATTCCGACTACACTGA

ref|AC_000155.1| TGGGCTTTATTCCCTTCCATTGATATATTCTTATAATAATGATAATTCCGACTACACTGA

************************************************************

ref|AC_000066.1| TCATTGTTAGTTTATAAGCCTTGAAATGAAGTAGGGTAACCCCACCACATTTGTTCTATT

ref|NC_000023.9| TCATTGTTAGTTTATAAGCCTTGAAATGAAGTAGGGTAACCCCACCACATTTGTTCTATT

ref|AC_000155.1| TCATTGTTAGTTTATAAGCCTTGAAATGAAGTAGGGTAACCCCACCACATTTGTTCTATT

************************************************************

ref|AC_000066.1| TTTTATTTATTTATTTATTTATTTATTTATTTATTTATTATTATTATACTTTAAGTTTTA

ref|NC_000023.9| TTTTATTTATTTATTTATTTATTTATTTATTTATTTATTATTATTATACTTTAAGTTTTA

ref|AC_000155.1| TTTTATTTATTTATTTATTTATTTATTTATTTATTTATTATTATTATACTTTAAGTTTTA

************************************************************

ref|AC_000066.1| GGGTACATGTGCACAATGTGCAGGTTAGTTACACATGTATACATGTGCCATGCTGGTGCG

ref|NC_000023.9| GGGTACATGTGCACAATGTGCAGGTTAGTTACACATGTATACATGTGCCATGCTGGTGCG

ref|AC_000155.1| GGGTACATGTGCACAATGTGCAGGTTAGTTACACATGTATACATGTGCCATGCTGGTGCG

************************************************************

ref|AC_000066.1| CTGCACCCACTAACTCGTCA

ref|NC_000023.9| CTGCACCCACTAACTCGTCA

ref|AC_000155.1| CTGCACCCACTAACTCGTCA

********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152155.2	NC_000023	152155298--152155326	14.5	100	58
	AC_000066	152661789--152661815	13.5	100	54
	AC_000155	141161800--141161826	13.5	100	54

HEMA152155.2	Information
Amplicon	260-262 bp
Forward ampliprimer	TGTTGTCATCCTGTGAGTCAA
Reverse ampliprimer	GCAAAGGATTCTTCCACCAG

>ref|NC_000023.9|

TGTTGTCATCCTGTGAGTCAATTATCCTTAATAAATTCCCTTTCTCATATATATATATATATATATAAAATATCTATACATTCATATATA

TACATATATCTATGAAATCTATAGATATATGAAATGTATACACACACACACACACACACACACACACATTTATCTTACTAGTTCTGTCCC

TCTAGAGAACTCTAATACACTACTTATGTTTCACTGTCACAAAACCTTTTGGTTCATATAAACTGGTGGAAGAATCCTTTGC

>ref|AC_000066.1|

TGTTGTCATCCTGTGAGTCAATTATCCTTAATAAATTCCCTTTCTCATATATATATATATATATATATAAAATATCTATACATTCATATA

TATACATATATCTATGAAATCTATAGATATATGAAATGTATACACACACACACACACACACACACACATTTATCTTACTAGTTCTGTCCC

TCTAGAGAACTCTAATACACTACTTATGTTTCACTGTCACAAAACCTTTTGGTTCATATAAACTGGTGGAAGAATCCTTTGC

>ref|AC_000155.1|

TGTTGTCATCCTGTGAGTCAATTATCCTTAATAAATTCCCTTTCTCATATATATATATATATATATAAAATATCTATACATTCATATATA

TACATATATCTATGAAATCTATAGATATATGAAATGTATACACACACACACACACACACACACACATTTATCTTACTAGTTCTGTCCCTC

TAGAGAACTCTAATACACTACTTATGTTTCACTGTCACAAAACCTTTTGGTTCATATAAACTGGTGGAAGAATCCTTTGC

HEMA152155.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 262 bp

Sequence 2: ref|AC_000066.1| 262 bp

Sequence 3: ref|AC_000155.1| 260 bp

ref|AC_000066.1| TGTTGTCATCCTGTGAGTCAATTATCCTTAATAAATTCCCTTTCTCATATATATATATAT

ref|AC_000155.1| TGTTGTCATCCTGTGAGTCAATTATCCTTAATAAATTCCCTTTCTCATATATATATATAT

ref|NC_000023.9| TGTTGTCATCCTGTGAGTCAATTATCCTTAATAAATTCCCTTTCTCATATATATATATAT

************************************************************

ref|AC_000066.1| ATATATATAAAATATCTATACATTCATATATATACATATATCTATGAAATCTATAGATAT

ref|AC_000155.1| ATATATA--AAATATCTATACATTCATATATATACATATATCTATGAAATCTATAGATAT

ref|NC_000023.9| ATATATA--AAATATCTATACATTCATATATATACATATATCTATGAAATCTATAGATAT

******* ***************************************************

ref|AC_000066.1| ATGAAATGTATACACACACACACACACACACACACACA--TTTATCTTACTAGTTCTGTC

ref|AC_000155.1| ATGAAATGTATACACACACACACACACACACACACACA--TTTATCTTACTAGTTCTGTC

ref|NC_000023.9| ATGAAATGTATACACACACACACACACACACACACACACATTTATCTTACTAGTTCTGTC

************************************** ********************

ref|AC_000066.1| CCTCTAGAGAACTCTAATACACTACTTATGTTTCACTGTCACAAAACCTTTTGGTTCATA

ref|AC_000155.1| CCTCTAGAGAACTCTAATACACTACTTATGTTTCACTGTCACAAAACCTTTTGGTTCATA

ref|NC_000023.9| CCTCTAGAGAACTCTAATACACTACTTATGTTTCACTGTCACAAAACCTTTTGGTTCATA

************************************************************

ref|AC_000066.1| TAAACTGGTGGAAGAATCCTTTGC

ref|AC_000155.1| TAAACTGGTGGAAGAATCCTTTGC

ref|NC_000023.9| TAAACTGGTGGAAGAATCCTTTGC

************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152155.6	NC_000023	152155655--152155695	20.5	89	64
	AC_000066	152661424--152661460	18.5	88	56
	AC_000155	141162155--141162191	18.5	88	56

HEMA152155.6	Information
Amplicon	193-197 bp
Forward ampliprimer	TGGATGTACATTTCCCATTTCC
Reverse ampliprimer	TCTGTTTGCCTGTGAAGCAC

>ref|NC_000023.9|

TGGATGTACATTTCCCATTTCCATACATTAGAAACACATAGGTCTTTTCTACTATGCATATATATGTATATATGTGTGTATATATATATG

TGTGTGTGTGTGTGTGTGTGTGTGTGTGCCTGTGTGTGTTTTGTGTACCATCATTTGTATTGGAAGGTAAGGTGTTATTGTTGAAGCGTG

CTTCACAGGCAAACAGA

>ref|AC_000066.1|

TGGATGTACATTTCCCATTTCCATACATTAGAAACACATAGGTCTTTTCTACTATGCATATATATGTATATATGTGTGTATATATATATG

TGTGTGTGTGTGTGTGTGTGTGTGCCTGTGTGTGTTTTGTGTACCATCATTTGTATTGGAAGGTAAGGTGTTATTGTTGAAGCGTGCTTC

ACAGGCAAACAGA

>ref|AC_000155.1|

TGGATGTACATTTCCCATTTCCATACATTAGAAACACATAGGTCTTTTCTACTATGCATATATATGTATATATGTGTGTATATATATATG

TGTGTGTGTGTGTGTGTGTGTGTGCCTGTGTGTGTTTTGTGTACCATCATTTGTATTGGAAGGTAAGGTGTTATTGTTGAAGCGTGCTTC

ACAGGCAAACAGA

HEMA152155.6

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 197 bp

Sequence 2: ref|AC_000066.1| 193 bp

Sequence 3: ref|AC_000155.1| 193 bp

ref|AC_000066.1| TGGATGTACATTTCCCATTTCCATACATTAGAAACACATAGGTCTTTTCTACTATGCATA

ref|AC_000155.1| TGGATGTACATTTCCCATTTCCATACATTAGAAACACATAGGTCTTTTCTACTATGCATA

ref|NC_000023.9| TGGATGTACATTTCCCATTTCCATACATTAGAAACACATAGGTCTTTTCTACTATGCATA

************************************************************

ref|AC_000066.1| TATATGTATATATGTGTGTATATATATATGTGTGTGTGTGTGTGTGTGTGTGTG----CC

ref|AC_000155.1| TATATGTATATATGTGTGTATATATATATGTGTGTGTGTGTGTGTGTGTGTGTG----CC

ref|NC_000023.9| TATATGTATATATGTGTGTATATATATATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGCC

****************************************************** **

ref|AC_000066.1| TGTGTGTGTTTTGTGTACCATCATTTGTATTGGAAGGTAAGGTGTTATTGTTGAAGCGTG

ref|AC_000155.1| TGTGTGTGTTTTGTGTACCATCATTTGTATTGGAAGGTAAGGTGTTATTGTTGAAGCGTG

ref|NC_000023.9| TGTGTGTGTTTTGTGTACCATCATTTGTATTGGAAGGTAAGGTGTTATTGTTGAAGCGTG

************************************************************

ref|AC_000066.1| CTTCACAGGCAAACAGA

ref|AC_000155.1| CTTCACAGGCAAACAGA

ref|NC_000023.9| CTTCACAGGCAAACAGA

*****************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**LOC649201Int2**	NC_000023	152250679--152250715	18.5	100	74
	AC_000066	152830452--152830488	18.5	100	74
	AC_000155	141256874--141256894	10.5	100	42

LOC649201Int2	Information
Amplicon	215-231 bp
Forward ampliprimer	TTGCCTTGATTGTGGTGATG
Reverse ampliprimer	GTTCACGTCTTTTGCCCATT

>ref|NC_000023.9|

TTGCCTTGATTGTGGTGATGGTTTCATACGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGAAAGCTTATCAAGTTGTTTAGCTG

AAACATGACAAATTCACAGTTTATTGTCTGTCAAATATATCTCAGTGTAAACAGACAACCTACAGAATGGGAGAAAATCACCTGACAAAG

GTCTAATATCCAGAATATAACCCCATCAAAAAATGGGCAAAAGACGTGAAC

>ref|AC_000066.1|

TTGCCTTGATTGTGGTGATGGTTTCATACGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGAAAGCTTATCAAGTTGTTTAGCTG

AAACATGACAAATTCACAGTTTATTGTCTGTCAAATATATCTCAGTGTAAACAGACAACCTACAGAATGGGAGAAAATCACCTGACAAAG

GTCTAATATCCAGAATATAACCCCATCAAAAAATGGGCAAAAGACGTGAAC

>ref|AC_000155.1|

TTGCCTTGATTGTGGTGATGGTTTCATACGTGTGTGTGTGTGTGTGTGTGAAAGCTTATCAAGTTGTTTAGCTGAAACATGACAAATTCA

CAGTTTATTGTCTGTCAAATATATCTCAGTGTAAACAGACAACCTACAGAATGGGAGAAAATCACCTGACAAAGGTCTAATATCCAGAAT

ATAACCCCATCAAAAAATGGGCAAAAGACGTGAAC

LOC649201Int2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 231 bp

Sequence 2: ref|AC_000066.1| 231 bp

Sequence 3: ref|AC_000155.1| 215 bp

ref|AC_000066.1| TTGCCTTGATTGTGGTGATGGTTTCATACGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|AC_000155.1| TTGCCTTGATTGTGGTGATGGTTTCATACGTGTGTGTGTGTGTGTGTGTG----------

ref|NC_000023.9| TTGCCTTGATTGTGGTGATGGTTTCATACGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

**************************************************

ref|AC_000066.1| TGTGTGAAAGCTTATCAAGTTGTTTAGCTGAAACATGACAAATTCACAGTTTATTGTCTG

ref|AC_000155.1| ------AAAGCTTATCAAGTTGTTTAGCTGAAACATGACAAATTCACAGTTTATTGTCTG

ref|NC_000023.9| TGTGTGAAAGCTTATCAAGTTGTTTAGCTGAAACATGACAAATTCACAGTTTATTGTCTG

******************************************************

ref|AC_000066.1| TCAAATATATCTCAGTGTAAACAGACAACCTACAGAATGGGAGAAAATCACCTGACAAAG

ref|AC_000155.1| TCAAATATATCTCAGTGTAAACAGACAACCTACAGAATGGGAGAAAATCACCTGACAAAG

ref|NC_000023.9| TCAAATATATCTCAGTGTAAACAGACAACCTACAGAATGGGAGAAAATCACCTGACAAAG

************************************************************

ref|AC_000066.1| GTCTAATATCCAGAATATAACCCCATCAAAAAATGGGCAAAAGACGTGAAC

ref|AC_000155.1| GTCTAATATCCAGAATATAACCCCATCAAAAAATGGGCAAAAGACGTGAAC

ref|NC_000023.9| GTCTAATATCCAGAATATAACCCCATCAAAAAATGGGCAAAAGACGTGAAC

***************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152257.3	NC_000023	152257313--152257369	28.5	100	114
	AC_000066	152837086--152837142	28.5	100	114
	AC_000155	141263464--141263506	21.5	100	86

HEMA152257.3	Information
Amplicon	246-260 bp
Forward ampliprimer	TTTCAACAGTTAATCATGCCAAT
Reverse ampliprimer	TGCTGCCTTTAAACCATCCT

>ref|NC_000023.9|

TTTCAACAGTTAATCATGCCAATTAAGTAAAAACACTTCAGTAGGATTTGCCCGATTAAGTTGAACACTTCAGGAGGATTCACTCAATTA

AATGTGGGTGGCTTGGGAGGACTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAAGAGACAGAG

AGAGACGAAGAAAGAGAGAGAGGAATGGCATGTCTCGTTGTGCAGTGAGGGCATGTTAACAGGATGGTTTAAAGGCAGCA

>ref|AC_000066.1|

TTTCAACAGTTAATCATGCCAATTAAGTAAAAACACTTCAGTAGGATTTGCCCGATTAAGTTGAACACTTCAGGAGGATTCACTCAATTA

AATGTGGGTGGCTTGGGAGGACTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAAGAGACAGAG

AGAGACGAAGAAAGAGAGAGAGGAATGGCATGTCTCGTTGTGCAGTGAGGGCATGTTAACAGGATGGTTTAAAGGCAGCA

>ref|AC_000155.1|

TTTCAACAGTTAATCATGCCAATTAAGTAAAAACACTTCAGTAGGATTTGCCCGATTAAGTTGAACACTTCAGGAGGATTCACTCAATTA

AATGTGGGTGGCTTGGGAGGACTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAAGAGACAGAGAGAGACGAAGAAAG

AGAGAGAGGAATGGCATGTCTCGTTGTGCAGTGAGGGCATGTTAACAGGATGGTTTAAAGGCAGCA

HEMA152257.3

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 260 bp

Sequence 2: ref|AC_000066.1| 260 bp

Sequence 3: ref|AC_000155.1| 246 bp

ref|AC_000066.1| TTTCAACAGTTAATCATGCCAATTAAGTAAAAACACTTCAGTAGGATTTGCCCGATTAAG

ref|NC_000023.9| TTTCAACAGTTAATCATGCCAATTAAGTAAAAACACTTCAGTAGGATTTGCCCGATTAAG

ref|AC_000155.1| TTTCAACAGTTAATCATGCCAATTAAGTAAAAACACTTCAGTAGGATTTGCCCGATTAAG

************************************************************

ref|AC_000066.1| TTGAACACTTCAGGAGGATTCACTCAATTAAATGTGGGTGGCTTGGGAGGACTGTGTGTG

ref|NC_000023.9| TTGAACACTTCAGGAGGATTCACTCAATTAAATGTGGGTGGCTTGGGAGGACTGTGTGTG

ref|AC_000155.1| TTGAACACTTCAGGAGGATTCACTCAATTAAATGTGGGTGGCTTGGGAGGACTGTGTGTG

************************************************************

ref|AC_000066.1| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAAGAGACAGAG

ref|NC_000023.9| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAAGAGACAGAG

ref|AC_000155.1| TGTGTGTGTGTGTGTGTGTGTGTGTGTG--------------TGTGTGTAAGAGACAGAG

**************************** ******************

ref|AC_000066.1| AGAGACGAAGAAAGAGAGAGAGGAATGGCATGTCTCGTTGTGCAGTGAGGGCATGTTAAC

ref|NC_000023.9| AGAGACGAAGAAAGAGAGAGAGGAATGGCATGTCTCGTTGTGCAGTGAGGGCATGTTAAC

ref|AC_000155.1| AGAGACGAAGAAAGAGAGAGAGGAATGGCATGTCTCGTTGTGCAGTGAGGGCATGTTAAC

************************************************************

ref|AC_000066.1| AGGATGGTTTAAAGGCAGCA

ref|NC_000023.9| AGGATGGTTTAAAGGCAGCA

ref|AC_000155.1| AGGATGGTTTAAAGGCAGCA

********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS15	NC_000023	152285381--152285422	21.0	100	84
DXS15	AC_000066	152865107--152865148	21.0	100	84

DXS15	Information
Amplicon	250 bp
Forward ampliprimer	CATACGCAATAAATAGCACATGGT
Reverse ampliprimer	AGAGAATGAAGAAATTAGGTGCTGA

>ref|NC_000023.9|

CATACGCAATAAATAGCACATGGTATAATGAACCTCCACGTACCCACCACCCAACTTCAACAATTATCCATTGACATTCTATGCATGTAC

ATGCACACACACACACACACACACACACACACACACACACACACATTCCTATTTTATACAAATGCTAGCATGCTACTCACACTGTGCTTT

ACTTTTCTTTCTTAAATTAACAATGTGTGTTGGAGATGGTTTATATCAGCACCTAATTTCTTCATTCTCT

>ref|AC_000066.1|

CATACGCAATAAATAGCACATGGTATAATGAACCTCCACGTACCCACCACCCAACTTCAACAATTATCCATTGACATTCTATGCATGTAC

ATGCACACACACACACACACACACACACACACACACACACACACATTCCTATTTTATACAAATGCTAGCATGCTACTCACACTGTGCTTT

ACTTTTCTTTCTTAAATTAACAATGTGTGTTGGAGATGGTTTATATCAGCACCTAATTTCTTCATTCTCT

DXS15

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 250 bp

Sequence 2: ref|AC_000066.1| 250 bp

ref|NC_000023.9| CATACGCAATAAATAGCACATGGTATAATGAACCTCCACGTACCCACCACCCAACTTCAA

ref|AC_000066.1| CATACGCAATAAATAGCACATGGTATAATGAACCTCCACGTACCCACCACCCAACTTCAA

************************************************************

ref|NC_000023.9| CAATTATCCATTGACATTCTATGCATGTACATGCACACACACACACACACACACACACAC

ref|AC_000066.1| CAATTATCCATTGACATTCTATGCATGTACATGCACACACACACACACACACACACACAC

************************************************************

ref|NC_000023.9| ACACACACACACACATTCCTATTTTATACAAATGCTAGCATGCTACTCACACTGTGCTTT

ref|AC_000066.1| ACACACACACACACATTCCTATTTTATACAAATGCTAGCATGCTACTCACACTGTGCTTT

************************************************************

ref|NC_000023.9| ACTTTTCTTTCTTAAATTAACAATGTGTGTTGGAGATGGTTTATATCAGCACCTAATTTC

ref|AC_000066.1| ACTTTTCTTTCTTAAATTAACAATGTGTGTTGGAGATGGTTTATATCAGCACCTAATTTC

************************************************************

ref|NC_000023.9| TTCATTCTCT

ref|AC_000066.1| TTCATTCTCT

**********

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152317.9	NC_000023	152317955--152317988	17.5	93	61
	AC_000066	152898482--152898515	17.5	93	61
	AC_000155	141324043--141324076	17.5	93	61

HEMA152317.9	Information
Amplicon	251 bp
Forward ampliprimer	TCCTGTCCTTCATTCCTGAT
Reverse ampliprimer	AACCACGTGTGTTAGCATGTG

>ref|NC_000023.9|

TCCTGTCCTTCATTCCTGATATTGATTAGTGTTCATTCTCTCTTGTGTGTTGTGTGTGTGTGTGTGTGTGTGTGTGTTGTTTTTTGTTTG

TTTGTTTGTTTTTTGAGACAGGGTCTCACTCTGTCACCCAGGCTGGAGTGCAGCAGCTTGATCTCGGCTCAGTGCAGCCTCGACCCCCTG

AGCTCAAGCGATCCTCCCACCTCAGCCTCCTGAGTAGCTGGGACTACCGGCACATGCTAACACACGTGGTT

>ref|AC_000066.1|

TCCTGTCCTTCATTCCTGATATTGATTAGTGTTCATTCTCTCTTGTGTGTTGTGTGTGTGTGTGTGTGTGTGTGTGTTGTTTTTTGTTTG

TTTGTTTGTTTTTTGAGACAGGGTCTCACTCTGTCACCCAGGCTGGAGTGCAGCAGCTCGATCTCGGCTCAGTGCAGCCTCGACCCCCTG

AGCTCAAGCGATCCTCCCACCTCAGCCTCCTGAGTAGCTGGGACTACCGGCACATGCTAACACACGTGGTT

>ref|AC_000155.1|

TCCTGTCCTTCATTCCTGATATTGATTAGTGTTCATTCTCTCTTGTGTGTTGTGTGTGTGTGTGTGTGTGTGTGTGTTGTTTTTTGTTTG

TTTGTTTGTTTTTTGAGACAGGGTCTCACTCTGTCACCCAGGCTGGAGTGCAGCAGCTCGATCTCGGCTCAGTGCAGCCTCGACCCCCTG

AGCTCAAGCGATCCTCCCACCTCAGCCTCCTGAGTAGCTGGGACTACCGGCACATGCTAACACACGTGGTT

HEMA152317.9

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 251 bp

Sequence 2: ref|AC_000066.1| 251 bp

Sequence 3: ref|AC_000155.1| 251 bp

ref|AC_000066.1| TCCTGTCCTTCATTCCTGATATTGATTAGTGTTCATTCTCTCTTGTGTGTTGTGTGTGTG

ref|NC_000023.9| TCCTGTCCTTCATTCCTGATATTGATTAGTGTTCATTCTCTCTTGTGTGTTGTGTGTGTG

ref|AC_000155.1| TCCTGTCCTTCATTCCTGATATTGATTAGTGTTCATTCTCTCTTGTGTGTTGTGTGTGTG

************************************************************

ref|AC_000066.1| TGTGTGTGTGTGTGTGTTGTTTTTTGTTTGTTTGTTTGTTTTTTGAGACAGGGTCTCACT

ref|NC_000023.9| TGTGTGTGTGTGTGTGTTGTTTTTTGTTTGTTTGTTTGTTTTTTGAGACAGGGTCTCACT

ref|AC_000155.1| TGTGTGTGTGTGTGTGTTGTTTTTTGTTTGTTTGTTTGTTTTTTGAGACAGGGTCTCACT

************************************************************

ref|AC_000066.1| CTGTCACCCAGGCTGGAGTGCAGCAGCTCGATCTCGGCTCAGTGCAGCCTCGACCCCCTG

ref|NC_000023.9| CTGTCACCCAGGCTGGAGTGCAGCAGCTTGATCTCGGCTCAGTGCAGCCTCGACCCCCTG

ref|AC_000155.1| CTGTCACCCAGGCTGGAGTGCAGCAGCTCGATCTCGGCTCAGTGCAGCCTCGACCCCCTG

**************************** *******************************

ref|AC_000066.1| AGCTCAAGCGATCCTCCCACCTCAGCCTCCTGAGTAGCTGGGACTACCGGCACATGCTAA

ref|NC_000023.9| AGCTCAAGCGATCCTCCCACCTCAGCCTCCTGAGTAGCTGGGACTACCGGCACATGCTAA

ref|AC_000155.1| AGCTCAAGCGATCCTCCCACCTCAGCCTCCTGAGTAGCTGGGACTACCGGCACATGCTAA

************************************************************

ref|AC_000066.1| CACACGTGGTT

ref|NC_000023.9| CACACGTGGTT

ref|AC_000155.1| CACACGTGGTT

***********

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152342.0	NC_000023	152342046--152342085	20.0	100	80
	AC_000066	152922575--152922614	20.0	100	80
	AC_000155	141348201--141348242	21.0	100	84

HEMA152342.0	Information
Amplicon	183-185 bp
Forward ampliprimer	GGAGAATGCTGCTTGGAGTG
Reverse ampliprimer	AGGATGGGGAATAGGGACAC

>ref|NC_000023.9|

GGAGAATGCTGCTTGGAGTGCCAGTCACTAGGACAACTCCAGAAATAATGTTATACACACACACACACACACACACACACACACACACAC

ACACGATATATATAGTTTTATGAAAACTTCTAGAGTTCTTGTAAGTTGCACTTATACCGTAGCCTCCTTCAGTGTGTCCCTATTCCCCAT

CCT

>ref|AC_000066.1|

GGAGAATGCTGCTTGGAGTGCCAGTCACTAGGACAACTCCAGAAATAATGTTATACACACACACACACACACACACACACACACACACAC

ACACGATATATATAGTTTTATGAAAACTTCTAGAGTTCTTGTAAGTTGCACTTATACCGTAGCCTCCTTCAGTGTGTCCCTATTCCCCAT

CCT

>ref|AC_000155.1|

GGAGAATGCTGCTTGGAGTGCCAGTCACTAGGACAACTCCAGAAATAATGTTATACACACACACACACACACACACACACACACACACAC

ACACACGATATATATAGTTTTATGAAAACTTCTAGAGTTCTTGTAAGTTGCACTTATACCGTAGCCTCCTTCAGTGTGTCCCTATTCCCC

ATCCT

HEMA152342.0

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 183 bp

Sequence 2: ref|AC_000066.1| 183 bp

Sequence 3: ref|AC_000155.1| 185 bp

ref|NC_000023.9| GGAGAATGCTGCTTGGAGTGCCAGTCACTAGGACAACTCCAGAAATAATGTTATACACAC

ref|AC_000066.1| GGAGAATGCTGCTTGGAGTGCCAGTCACTAGGACAACTCCAGAAATAATGTTATACACAC

ref|AC_000155.1| GGAGAATGCTGCTTGGAGTGCCAGTCACTAGGACAACTCCAGAAATAATGTTATACACAC

************************************************************

ref|NC_000023.9| ACACACACACACACACACACACACACACACACAC--GATATATATAGTTTTATGAAAACT

ref|AC_000066.1| ACACACACACACACACACACACACACACACACAC--GATATATATAGTTTTATGAAAACT

ref|AC_000155.1| ACACACACACACACACACACACACACACACACACACGATATATATAGTTTTATGAAAACT

********************************** ************************

ref|NC_000023.9| TCTAGAGTTCTTGTAAGTTGCACTTATACCGTAGCCTCCTTCAGTGTGTCCCTATTCCCC

ref|AC_000066.1| TCTAGAGTTCTTGTAAGTTGCACTTATACCGTAGCCTCCTTCAGTGTGTCCCTATTCCCC

ref|AC_000155.1| TCTAGAGTTCTTGTAAGTTGCACTTATACCGTAGCCTCCTTCAGTGTGTCCCTATTCCCC

************************************************************

ref|NC_000023.9| ATCCT

ref|AC_000066.1| ATCCT

ref|AC_000155.1| ATCCT

*****

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS1356	NC_000023	152351238--152351282	11.3	90	72
	NC_000023	152351280--152351343	16.0	100	128
	AC_000066	152931768--152931812	11.3	90	72
	AC_000066	152931810--152931873	16.0	100	128

DXS1356	Information
Amplicon	286 bp
Forward ampliprimer	TTCAGTGAGCCAAGATCACG
Reverse ampliprimer	GAGGGGTCTTCTGGCTCTTC

>ref|NC_000023.9|

TTCAGTGAGCCAAGATCACGCCACTGCACTCCAGCCTGGGCGACACAGCAAGACTCCATCAAAAAAAAGAAAAAAAGGAGAGAGGGAGAG

AAAAAGAGAGGAAGGAAGGGAGAGAGGGAGGGAGAGAGGGAGGGAGGGAGGGAGGGAGGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAA

GGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAAAAGCACCACAAACCAGGTGGCTTAAAACACCAGAAATTTACTCTCTTATTGTTGAAG

AGCCAGAAGACCCCTC

>ref|AC_000066.1|

TTCAGTGAGCCAAGATCACGCCACTGCACTCCAGCCTGGGCGACAGAGCAAGACTCCATCAAAAAAAAGAAAAAAAGGAGAGAGGGAGAG

AAAAAGAGAGGAAGGAAGGGAGAGAGGGAGGGAGAGAGGGAGGGAGGGAGGGAGGGAGGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAA

GGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAAAAGCACCACAAACCAGGTGGCTTAAAACACCAGAAATTTACTCTCTTATTGTTGAAG

AGCCAGAAGACCCCTC

DXS1356

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 286 bp

Sequence 2: ref|AC_000066.1| 286 bp

ref|NC_000023.9| TTCAGTGAGCCAAGATCACGCCACTGCACTCCAGCCTGGGCGACACAGCAAGACTCCATC

ref|AC_000066.1| TTCAGTGAGCCAAGATCACGCCACTGCACTCCAGCCTGGGCGACAGAGCAAGACTCCATC

********************************************* **************

ref|NC_000023.9| AAAAAAAAGAAAAAAAGGAGAGAGGGAGAGAAAAAGAGAGGAAGGAAGGGAGAGAGGGAG

ref|AC_000066.1| AAAAAAAAGAAAAAAAGGAGAGAGGGAGAGAAAAAGAGAGGAAGGAAGGGAGAGAGGGAG

************************************************************

ref|NC_000023.9| GGAGAGAGGGAGGGAGGGAGGGAGGGAGGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAA

ref|AC_000066.1| GGAGAGAGGGAGGGAGGGAGGGAGGGAGGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAA

************************************************************

ref|NC_000023.9| GGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAAAAGCACCACAAACCAGGTGGCTTAAAA

ref|AC_000066.1| GGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAAAAGCACCACAAACCAGGTGGCTTAAAA

************************************************************

ref|NC_000023.9| CACCAGAAATTTACTCTCTTATTGTTGAAGAGCCAGAAGACCCCTC

ref|AC_000066.1| CACCAGAAATTTACTCTCTTATTGTTGAAGAGCCAGAAGACCCCTC

**********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**UCHL5IPInt8**	NC_000023	152373164--152373200	9.3	100	74
**UCHL5IPInt8**	AC_000066	152953680--152953716	9.3	100	74

UCHL5IPInt8	Information
Amplicon	168 bp
Forward ampliprimer	TTGCTGCTTCTTCACGGTCT
Reverse ampliprimer	GGGTGGGAAGCAGGAGAT

>ref|NC_000023.9|

TTGCTGCTTCTTCACGGTCTCCACGGCCTTCGCAGAGGTGTCAGCAACTGCCATGACCACTTGCAGCAGCTGGGGAGGGAGGGAGGGAGG

GAGGGAGGGAGGGAGGGAGCGAGCAGGCACTAAATGCCGCTGGTCTCACGTAGCGACGGCATCTCCTGCTTCCCACCC

>ref|AC_000066.1|

TTGCTGCTTCTTCACGGTCTCCACGGCCTTCGCAGAGGTGTCAGCAACTGCCATGACCACTTGCAGCAGCTGGGGAGGGAGGGAGGGAGG

GAGGGAGGGAGGGAGGGAGCGAGCAGGCACTAAATGCCGCTGGTCTCACGTAGCGACGGCATCTCCTGCTTCCCACCC

UCHL5IPInt8

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 168 bp

Sequence 2: ref|AC_000066.1| 168 bp

ref|NC_000023.9| TTGCTGCTTCTTCACGGTCTCCACGGCCTTCGCAGAGGTGTCAGCAACTGCCATGACCAC

ref|AC_000066.1| TTGCTGCTTCTTCACGGTCTCCACGGCCTTCGCAGAGGTGTCAGCAACTGCCATGACCAC

************************************************************

ref|NC_000023.9| TTGCAGCAGCTGGGGAGGGAGGGAGGGAGGGAGGGAGGGAGGGAGGGAGCGAGCAGGCAC

ref|AC_000066.1| TTGCAGCAGCTGGGGAGGGAGGGAGGGAGGGAGGGAGGGAGGGAGGGAGCGAGCAGGCAC

************************************************************

ref|NC_000023.9| TAAATGCCGCTGGTCTCACGTAGCGACGGCATCTCCTGCTTCCCACCC

ref|AC_000066.1| TAAATGCCGCTGGTCTCACGTAGCGACGGCATCTCCTGCTTCCCACCC

************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS9901(BGNExon8)	NC_000023	152427681--152427710	15.0	100	60
	NC_000023	152427714--152427758	22.5	100	90
	AC_000066	153008186--153008213	14.0	92	47
	AC_000066	153008217--153008261	22.5	100	90
	AC_000155	141432840--141432869	15.0	100	60
	AC_000155	141432873--141432917	22.5	100	90

*DXS9901(BGNExon8)*	Information
Amplicon	250-252 bp
Forward ampliprimer	CTTCGACCAGTCCTCCCTTC
Reverse ampliprimer	AGAAAGCAGTCCCAGCTCAA

>ref|NC_000023.9|

CTTCGACCAGTCCTCCCTTCTGTTCTCTCTTTCCCCGTCCTTCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTGTGTGTGTGTGTG

TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTTGTGCTTCCTCAGACCTTTCTCGCTTCTGAGCTTGGTGGCCTGTTCCCTCCATCTCT

CCGAACCTGGCTTCGCCTGTCCCTTTCACTCCACACCCTCTGGCCTTCTGCCTTGAGCTGGGACTGCTTTCT

>ref|AC_000066.1|

CTTCGACCAGTCCTCCCTTCTGTTCTCTCTTTCCCCGTCCTTCCTCTCTCTCTCTCTCTCTCTTTCTCTCTTTCTGTGTGTGTGTGTGTG

TGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTTGTGCTTCCTCAGACCTTTCTCGCTTCTGAGCTTGGTGGCCTGTTCCCTCCATCTCTCC

GAACCTGGCTTCGCCTGTCCCTTTCACTCCACACCCTCTGGCCTTCTGCCTTGAGCTGGGACTGCTTTCT

>ref|AC_000155.1|

CTTCGACCAGTCCTCCCTTCTGTTCTCTCTTTCCCCGTCCTTCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTGTGTGTGTGTGTG

TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTTGTGCTTCCTCAGACCTTTCTCGCTTCTGAGCTTGGTGGCCTGTTCCCTCCATCTCT

CCGAACCTGGCTTCGCCTGTCCCTTTCACTCCACACCCTCTGGCCTTCTGCCTTGAGCTGGGACTGCTTTCT

DXS9901(BGNExon8)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 252 bp

Sequence 2: ref|AC_000066.1| 250 bp

Sequence 3: ref|AC_000155.1| 252 bp

ref|NC_000023.9| CTTCGACCAGTCCTCCCTTCTGTTCTCTCTTTCCCCGTCCTTCCTCTCTCTCTCTCTCTC

ref|AC_000155.1| CTTCGACCAGTCCTCCCTTCTGTTCTCTCTTTCCCCGTCCTTCCTCTCTCTCTCTCTCTC

ref|AC_000066.1| CTTCGACCAGTCCTCCCTTCTGTTCTCTCTTTCCCCGTCCTTCCTCTCTCTCTCTCTCTC

************************************************************

ref|NC_000023.9| TCTCTCTCTCTCTTTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|AC_000155.1| TCTCTCTCTCTCTTTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|AC_000066.1| TCTTTCTCTC--TTTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

*** ****** ************************************************

ref|NC_000023.9| TCTTGTGCTTCCTCAGACCTTTCTCGCTTCTGAGCTTGGTGGCCTGTTCCCTCCATCTCT

ref|AC_000155.1| TCTTGTGCTTCCTCAGACCTTTCTCGCTTCTGAGCTTGGTGGCCTGTTCCCTCCATCTCT

ref|AC_000066.1| TCTTGTGCTTCCTCAGACCTTTCTCGCTTCTGAGCTTGGTGGCCTGTTCCCTCCATCTCT

************************************************************

ref|NC_000023.9| CCGAACCTGGCTTCGCCTGTCCCTTTCACTCCACACCCTCTGGCCTTCTGCCTTGAGCTG

ref|AC_000155.1| CCGAACCTGGCTTCGCCTGTCCCTTTCACTCCACACCCTCTGGCCTTCTGCCTTGAGCTG

ref|AC_000066.1| CCGAACCTGGCTTCGCCTGTCCCTTTCACTCCACACCCTCTGGCCTTCTGCCTTGAGCTG

************************************************************

ref|NC_000023.9| GGACTGCTTTCT

ref|AC_000155.1| GGACTGCTTTCT

ref|AC_000066.1| GGACTGCTTTCT

                      ************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152442.8	NC_000023	152442878--152442911	17.0	93	59
	AC_000066	153023367--153023400	17.0	93	59
	AC_000155	141448135--141448168	17.0	93	59

HEMA152442.8	Information
Amplicon	164 bp
Forward ampliprimer	GGCTTAGAAAATCCCTTTCG
Reverse ampliprimer	GAAAAAGCTTGCTAATCCCTAAA

>ref|NC_000023.9|

GGCTTAGAAAATCCCTTTCGGAAATTTGAGAGCTGTTCATTTCTGAGTTTTCCATGTGTCTTCGTGTGTGTGTGTCTGTGTGTGTGTGTG

TGTGTGTATTGGGGTGGTGTAGTTGCATTTTTAAGATACTTAACTCTTTAATTTAGGGATTAGCAAGCTTTTTC

>ref|AC_000066.1|

GGCTTAGAAAATCCCTTTCGGAAATTTGAGAGCTGTTCATTTCTGAGTTTTCCATGTGTCTTCGTGTGTGTGTGTCTGTGTGTGTGTGTG

TGTGTGTATTGGGGTGGTGTAGTTGCATTTTTAAGATACTTAACTCTTTAATTTAGGGATTAGCAAGCTTTTTC

>ref|AC_000155.1|

GGCTTAGAAAATCCCTTTCGGAAATTTGAGAGCTGTTCATTTCTGAGTTTTCCATGTGTCTTCGTGTGTGTGTGTCTGTGTGTGTGTGTG

TGTGTGTATTGGGGTGGTGTAGTTGCATTTTTAAGATACTTAACTCTTTAATTTAGGGATTAGCAAGCTTTTTC

HEMA152442.8

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 164 bp

Sequence 2: ref|AC_000066.1| 164 bp

Sequence 3: ref|AC_000155.1| 164 bp

ref|AC_000066.1| GGCTTAGAAAATCCCTTTCGGAAATTTGAGAGCTGTTCATTTCTGAGTTTTCCATGTGTC

ref|NC_000023.9| GGCTTAGAAAATCCCTTTCGGAAATTTGAGAGCTGTTCATTTCTGAGTTTTCCATGTGTC

ref|AC_000155.1| GGCTTAGAAAATCCCTTTCGGAAATTTGAGAGCTGTTCATTTCTGAGTTTTCCATGTGTC

************************************************************

ref|AC_000066.1| TTCGTGTGTGTGTGTCTGTGTGTGTGTGTGTGTGTGTATTGGGGTGGTGTAGTTGCATTT

ref|NC_000023.9| TTCGTGTGTGTGTGTCTGTGTGTGTGTGTGTGTGTGTATTGGGGTGGTGTAGTTGCATTT

ref|AC_000155.1| TTCGTGTGTGTGTGTCTGTGTGTGTGTGTGTGTGTGTATTGGGGTGGTGTAGTTGCATTT

************************************************************

ref|AC_000066.1| TTAAGATACTTAACTCTTTAATTTAGGGATTAGCAAGCTTTTTC

ref|NC_000023.9| TTAAGATACTTAACTCTTTAATTTAGGGATTAGCAAGCTTTTTC

ref|AC_000155.1| TTAAGATACTTAACTCTTTAATTTAGGGATTAGCAAGCTTTTTC

********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**ATP2B3Int20**	NC_000023	152489656--152489691	18.0	100	72
	AC_000066	153070144--153070179	18.0	100	72
	AC_000155	141494798--141494839	21.0	100	84

ATP2B3Int20	Information
Amplicon	251-257 bp
Forward ampliprimer	CCCTCCTCTTGCATAAGCTG
Reverse ampliprimer	ACTGCTTTTCAGCCCAACTG

>ref|NC_000023.9|

CCCTCCTCTTGCATAAGCTGTCATTTTGGGCCACTCTGTCCTTTAAGGAAGGGAGGAGGCCCATCCTGTCCCCCTTGGGAACAGAGCACG

TGCGCGCGTACACACACACACACACACACACACACACACACACACTCCTTCTCCAAACCACAGGCCACGTACACCTCCAGCTTGCTCCAC

ACTCATCATGGGGCCACCTACCCTTGTCACAAATATTGGCTGACACCGTGGCAGTTGGGCTGAAAAGCAGT

>ref|AC_000066.1|

CCCTCCTCTTGCATAAGCTGTCATTTTGGGCCACTCTGTCCTTTAAGGAAGGGAGGAGGCCCATCCTGTCCCCCTTGGGAACAGAGCACG

TGCGCGCGTACACACACACACACACACACACACACACACACACACTCCTTCTCCAAACCACAGGCCACGTACACCTCCAGCTTGCTCCAC

ACTCATCATGGGGCCACCTACCCTTGTCACAAATATTGGCTGACACCGTGGCAGTTGGGCTGAAAAGCAGT

>ref|AC_000155.1|

CCCTCCTCTTGCATAAGCTGTCATTTTGGGCCACTCTGTCCTTTAAGGAAGGGAGGAGGCCCATCCTGTCCCCCTTGGGAACAGAGCACG

TGCGCGCGTACACACACACACACACACACACACACACACACACACACACACTCCTTCTCCAAACCACAGGCCACGTACACCTCCAGCTTG

CTCCACACTCATCATGGGGCCACCTACCCTTGTCACAAATATTGGCTGACACCGTGGCAGTTGGGCTGAAAAGCAGT

ATP2B3Int20

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 251 bp

Sequence 2: ref|AC_000066.1| 251 bp

Sequence 3: ref|AC_000155.1| 257 bp

ref|NC_000023.9| CCCTCCTCTTGCATAAGCTGTCATTTTGGGCCACTCTGTCCTTTAAGGAAGGGAGGAGGC

ref|AC_000066.1| CCCTCCTCTTGCATAAGCTGTCATTTTGGGCCACTCTGTCCTTTAAGGAAGGGAGGAGGC

ref|AC_000155.1| CCCTCCTCTTGCATAAGCTGTCATTTTGGGCCACTCTGTCCTTTAAGGAAGGGAGGAGGC

************************************************************

ref|NC_000023.9| CCATCCTGTCCCCCTTGGGAACAGAGCACGTGCGCGCGTACACACACACACACACACACA

ref|AC_000066.1| CCATCCTGTCCCCCTTGGGAACAGAGCACGTGCGCGCGTACACACACACACACACACACA

ref|AC_000155.1| CCATCCTGTCCCCCTTGGGAACAGAGCACGTGCGCGCGTACACACACACACACACACACA

************************************************************

ref|NC_000023.9| CACACACACACACAC------TCCTTCTCCAAACCACAGGCCACGTACACCTCCAGCTTG

ref|AC_000066.1| CACACACACACACAC------TCCTTCTCCAAACCACAGGCCACGTACACCTCCAGCTTG

ref|AC_000155.1| CACACACACACACACACACACTCCTTCTCCAAACCACAGGCCACGTACACCTCCAGCTTG

*************** ***************************************

ref|NC_000023.9| CTCCACACTCATCATGGGGCCACCTACCCTTGTCACAAATATTGGCTGACACCGTGGCAG

ref|AC_000066.1| CTCCACACTCATCATGGGGCCACCTACCCTTGTCACAAATATTGGCTGACACCGTGGCAG

ref|AC_000155.1| CTCCACACTCATCATGGGGCCACCTACCCTTGTCACAAATATTGGCTGACACCGTGGCAG

************************************************************

ref|NC_000023.9| TTGGGCTGAAAAGCAGT

ref|AC_000066.1| TTGGGCTGAAAAGCAGT

ref|AC_000155.1| TTGGGCTGAAAAGCAGT

*****************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152503.2	NC_000023	152503201--152503236	9.0	100	72
	AC_000066	153083688--153083727	10.0	100	80
	AC_000155	141508566--141508605	10.0	100	80

HEMA152503.2	Information
Amplicon	234-254 bp
Forward ampliprimer	CTCCAGCCTGAGTGACAGAG
Reverse ampliprimer	ACACCAATGCAGCAAGGAC

>ref|NC_000023.9|

CTCCAGCCTGAGTGACAGAGTAAGATTCTGTCAGGAAAGAAAAGAAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGGAGGGAGGG

AAGAGGAGAGGAGAGGAGAGAAGACAAGAGAAGAGAAGAATAAGAGGAGAAGGAAAAGAAAAGAAAAAAGAGAAAAGAGAAGAGAAAAGA

CAAATCAAAAACAGCAGGCAGAAGAATCGCAGCATGTCCTTGCTGCATTGGTGT

>ref|AC_000066.1|

CTCCAGCCTGAGTGACAGAGTAAGATTCTGTCAGGAAAGAAAAGAAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGGAGGG

AAGAGGAGAGGAGAGGAGAGAAGACAAGAGAAGAGAAGAATAAGAGGAGAAGGAAAAGAAAAGAAAAAAGAGAAAAGAGAAGAGAAAAGA

CAAATCAAAAACAGCAGGCAGAAGAATCGCAGCATGTCCTTGCTGCATTGGTGT

>ref|AC_000155.1|

CTCCAGCCTGAGTGACAGAGTAAGATTCTGTCAGGAAAGAAAAGAAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGGAGGG

AGGGAGGGAGGGAGGGAGGGAAGAGGAGAGGAGAGGAGAGAAGACAAGAGAAGAGAAGAATAAGAGGAGAAGGAAAAGAAAAGAAAAAAG

AGAAAAGAGAAGAGAAAAGACAAATCAAAAACAGCAGGCAGAAGAATCGCAGCATGTCCTTGCTGCATTGGTGT

HEMA152503.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 234 bp

Sequence 2: ref|AC_000066.1| 234 bp

Sequence 3: ref|AC_000155.1| 254 bp

ref|NC_000023.9| CTCCAGCCTGAGTGACAGAGTAAGATTCTGTCAGGAAAGAAAAGAAAGGAAGGAAGGAAG

ref|AC_000066.1| CTCCAGCCTGAGTGACAGAGTAAGATTCTGTCAGGAAAGAAAAGAAAGGAAGGAAGGAAG

ref|AC_000155.1| CTCCAGCCTGAGTGACAGAGTAAGATTCTGTCAGGAAAGAAAAGAAAGGAAGGAAGGAAG

************************************************************

ref|NC_000023.9| GAAGGAAGGAAGGAAGGAAGG--------------------GAGGGAGGGAAGAGGAGAG

ref|AC_000066.1| GAAGGAAGGAAGGAAGGAAGG--------------------AAGGGAGGGAAGAGGAGAG

ref|AC_000155.1| GAAGGAAGGAAGGAAGGAAGGAAGGGAGGGAGGGAGGGAGGGAGGGAGGGAAGAGGAGAG

********************* ******************

ref|NC_000023.9| GAGAGGAGAGAAGACAAGAGAAGAGAAGAATAAGAGGAGAAGGAAAAGAAAAGAAAAAAG

ref|AC_000066.1| GAGAGGAGAGAAGACAAGAGAAGAGAAGAATAAGAGGAGAAGGAAAAGAAAAGAAAAAAG

ref|AC_000155.1| GAGAGGAGAGAAGACAAGAGAAGAGAAGAATAAGAGGAGAAGGAAAAGAAAAGAAAAAAG

************************************************************

ref|NC_000023.9| AGAAAAGAGAAGAGAAAAGACAAATCAAAAACAGCAGGCAGAAGAATCGCAGCATGTCCT

ref|AC_000066.1| AGAAAAGAGAAGAGAAAAGACAAATCAAAAACAGCAGGCAGAAGAATCGCAGCATGTCCT

ref|AC_000155.1| AGAAAAGAGAAGAGAAAAGACAAATCAAAAACAGCAGGCAGAAGAATCGCAGCATGTCCT

************************************************************

ref|NC_000023.9| TGCTGCATTGGTGT

ref|AC_000066.1| TGCTGCATTGGTGT

ref|AC_000155.1| TGCTGCATTGGTGT

**************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152520.9	NC_000023	152520968--152521000	8.3	100	66
	AC_000066	153101469--153101501	8.3	100	66
	AC_000155	141526532--141526564	8.3	100	66

HEMA152520.9	Information
Amplicon	179 bp
Forward ampliprimer	ATCCACAGCCTCCATCTCTG
Reverse ampliprimer	GGGAGGTGGAGGTTACAGTG

>ref|NC_000023.9|

ATCCACAGCCTCCATCTCTGTTCTCCCTCCCCGCTCCCCACTGCCTACCCACCCTTCATCTTTCTATTCAATTATTTATTTATTTATTTA

TTTATTTATTTATTGAGACAGATT CTCACTCTGCCGCCCAGGCTGGAGTGCAGTAGCATGATCTTGGCTCACTGTAACCTCCACCTCCC

>ref|AC_000066.1|

ATCCACAGCCTCCATCTCTGTTCTCCCTCCCCGCTCCCCACTGCCTACCCACCCTTCATCTTTCTATTCAATTATTTATTTATTTATTTA

TTTATTTATTTATTGAGACAGATTCTCACTCTGCCGCCCAGGCTGGAGTGCAGTAGCATGATCTTGGCTCACTGTAACCTCCACCTCCC

>ref|AC_000155.1|

ATCCACAGCCTCCATCTCTGTTCTCCCTCCCCGCTCCCCACTGCCTACCCACCCTTCATCTTTCTATTCAATTATTTATTTATTTATTTA

TTTATTTATTTATTGAGACAGATTCTCACTCTGCCGCCCAGGCTGGAGTGCAGTAGCATGATCTTGGCTCACTGTAACCTCCACCTCCC

HEMA152520.9

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 179 bp

Sequence 2: ref|AC_000066.1| 179 bp

Sequence 3: ref|AC_000155.1| 179 bp

ref|NC_000023.9| ATCCACAGCCTCCATCTCTGTTCTCCCTCCCCGCTCCCCACTGCCTACCCACCCTTCATC

ref|AC_000155.1| ATCCACAGCCTCCATCTCTGTTCTCCCTCCCCGCTCCCCACTGCCTACCCACCCTTCATC

ref|AC_000066.1| ATCCACAGCCTCCATCTCTGTTCTCCCTCCCCGCTCCCCACTGCCTACCCACCCTTCATC

************************************************************

ref|NC_000023.9| TTTCTATTCAATTATTTATTTATTTATTTATTTATTTATTTATTGAGACAGATTCTCACT

ref|AC_000155.1| TTTCTATTCAATTATTTATTTATTTATTTATTTATTTATTTATTGAGACAGATTCTCACT

ref|AC_000066.1| TTTCTATTCAATTATTTATTTATTTATTTATTTATTTATTTATTGAGACAGATTCTCACT

************************************************************

ref|NC_000023.9| CTGCCGCCCAGGCTGGAGTGCAGTAGCATGATCTTGGCTCACTGTAACCTCCACCTCCC

ref|AC_000155.1| CTGCCGCCCAGGCTGGAGTGCAGTAGCATGATCTTGGCTCACTGTAACCTCCACCTCCC

ref|AC_000066.1| CTGCCGCCCAGGCTGGAGTGCAGTAGCATGATCTTGGCTCACTGTAACCTCCACCTCCC

***********************************************************

Locus name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152524.1	NC_000023	152524121--152524353	56.5	88	285
	AC_000066	153104622--153104850	55.5	88	284
	AC_000155	141528559--141528756	48.5	87	251

HEMA152524.1	Information
Amplicon	257-292 bp
Forward ampliprimer	AAAATCCCAGCTGGCTTAAA
Reverse ampliprimer	TCAGCTTGTCAATGTCTGCAC

>ref|NC_000023.9|

AAAATCCCAGCTGGCTTAAATTAAGAAAGAAAGAAAGAAAAATAAAGAAGAAAGAAAGAAAGAGAGAGAGAGAGAAAGAAAGGAAAGGAA

AGAAAGAAAAGAAGGAAAGAAAGGAAAGAAAGAAAGAAAGGAAAGAAAGAAAGAAAGAAGGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA

AAGAAAGAAAGAAAGAAAGAAAGAAAAAAGAAAGAAAGAAAAGAAAAGAAAAGAAAAGAAAAGAAAGAAAAGAAAATCCTAGCTGGCTTT

TGTGCAGACATTGACAAGCTGA

>ref|AC_000066.1|

AAAATCCCAGCTGGCTTAAATTAAGAAAGAAAGAAAGAAAAATAAAGAAGAAAGAAAGAAAGAGAGAGAGAGAGAAAGAAAGGAAAGGAA

AGAAAGAAAAGAAGGAAAGAAAGGAAAGAAAGAAAGAAAGGAAAGAAAGAAAGAAAGAAGGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA

AAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAAAAAGAAAAGAAAAGAAAAGAAAAGAAAGAAAAGAAAATCCTAGCTGGCTTTTGTG

CAGACATTGACAAGCTGA

>ref|AC_000155.1|

AAAATCCCAGCTGGCTTAAATTAAGAAAGAAAGAAAGAAAAATAAAGAAGAAAGAAAGAAAGAGAGAGAGAGAGAAAGAAAGGAAAGGAA

AGAAAGAAAAGAAGGAAAGAAAGGAAAGAAAGAAAGAAAGAAAGGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA

AAGAAAAAAGAAAGAAAGAAAAGAAAAGAAAGAAAAGAAAATCCTAGCTGGCTTTTGTGCAGACATTGACAAGCTGA

HEMA152524.1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 292 bp

Sequence 2: ref|AC_000066.1| 288 bp

Sequence 3: ref|AC_000155.1| 257 bp

ref|NC_000023.9| AAAATCCCAGCTGGCTTAAATTAAGAAAGAAAGAAAGAAAAATAAAGAAGAAAGAAAGAA

ref|AC_000066.1| AAAATCCCAGCTGGCTTAAATTAAGAAAGAAAGAAAGAAAAATAAAGAAGAAAGAAAGAA

ref|AC_000155.1| AAAATCCCAGCTGGCTTAAATTAAGAAAGAAAGAAAGAAAAATAAAGAAGAAAGAAAGAA

************************************************************

ref|NC_000023.9| AGAGAGAGAGAGAGAAAGAAAGGAAAGGAAAGAAAGAAAAGAAGGAAAGAAAGGAAAGAA

ref|AC_000066.1| AGAGAGAGAGAGAGAAAGAAAGGAAAGGAAAGAAAGAAAAGAAGGAAAGAAAGGAAAGAA

ref|AC_000155.1| AGAGAGAGAGAGAGAAAGAAAGGAAAGGAAAGAAAGAAAAGAAGGAAAGAAAGGAAAGAA

************************************************************

ref|NC_000023.9| AGAAAGAAAGGAAAGAAAGAAAGAAAGAAGGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA

ref|AC_000066.1| AGAAAGAAAGGAAAGAAAGAAAGAAAGAAGGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA

ref|AC_000155.1| AGAAAGAAAGAAAGGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA

********** ** *************** ******************************

ref|NC_000023.9| AAGAAAGAAAGAAAGAAAGAAAGAAAAAAGAAAGAAAGAAAAGAAAAGAAAAGAAAAGAA

ref|AC_000066.1| AAGAAAGAAAGAAAGAAAGAAAGAAA---GAAAGAAA-AAAAGAAAAGAAAAGAAAAGAA

ref|AC_000155.1| AAGAAA-----AAAGAAAGAAAGAAA------------------------------AGAA

****** *************** ****

ref|NC_000023.9| AAGAAAGAAAAGAAAATCCTAGCTGGCTTTTGTGCAGACATTGACAAGCTGA

ref|AC_000066.1| AAGAAAGAAAAGAAAATCCTAGCTGGCTTTTGTGCAGACATTGACAAGCTGA

ref|AC_000155.1| AAGAAAGAAAAGAAAATCCTAGCTGGCTTTTGTGCAGACATTGACAAGCTGA

****************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152539.8	NC_000023	152539891--152539990	20.2	89	112
HEMA152539.8	AC_000066	153120380--153120479	20.2	89	112

HEMA152539.8	Information
Amplicon	254 bp
Forward ampliprimer	GGTTGTGGTGAGCCAAGATT
Reverse ampliprimer	TGGGTGACAGAGCAAGACTG

>ref|NC_000023.9|

GGTTGTGGTGAGCCAAGATTGCACCACTGCACTCCAGCCTAGGCGACAGAGCAAGACTCCGTCTCAGAAAAAAAAAAAAAAGAAAAGAAA

AGGAAAGAAAAGAAAGGAAAGGAAAAGAAAAGAAAGGAAAGGAAAGGAAAGGAAAAGAAAGAAAAGAAAAGAAAAAGACAAGACAGGAAA

AGCAGTATTTTTATTGTACACACCTGGTGGGATATATGTGTTTTTTTTGAAACACAGTCTTGCTCTGTCACCCA

>ref|AC_000066.1|

GGTTGTGGTGAGCCAAGATTGCACCACTGCACTCCAGCCTAGGCGACAGAGCAAGACTCCGTCTCAGAAAAAAAAAAAAAAGAAAAGAAA

AGGAAAGAAAAGAAAGGAAAGGAAAAGAAAAGAAAGGAAAGGAAAGGAAAGGAAAAGAAAGAAAAGAAAAGAAAAAGACAAGACAGGAAA

AGCAGTATTTTTATTGTACACACCTGGTGGGATATATGTGTTTTTTTTGAAACACAGTCTTGCTCTGTCACCCA

HEMA152539.8

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 254 bp

Sequence 2: ref|AC_000066.1| 254 bp

ref|NC_000023.9| GGTTGTGGTGAGCCAAGATTGCACCACTGCACTCCAGCCTAGGCGACAGAGCAAGACTCC

ref|AC_000066.1| GGTTGTGGTGAGCCAAGATTGCACCACTGCACTCCAGCCTAGGCGACAGAGCAAGACTCC

************************************************************

ref|NC_000023.9| GTCTCAGAAAAAAAAAAAAAAGAAAAGAAAAGGAAAGAAAAGAAAGGAAAGGAAAAGAAA

ref|AC_000066.1| GTCTCAGAAAAAAAAAAAAAAGAAAAGAAAAGGAAAGAAAAGAAAGGAAAGGAAAAGAAA

************************************************************

ref|NC_000023.9| AGAAAGGAAAGGAAAGGAAAGGAAAAGAAAGAAAAGAAAAGAAAAAGACAAGACAGGAAA

ref|AC_000066.1| AGAAAGGAAAGGAAAGGAAAGGAAAAGAAAGAAAAGAAAAGAAAAAGACAAGACAGGAAA

************************************************************

ref|NC_000023.9| AGCAGTATTTTTATTGTACACACCTGGTGGGATATATGTGTTTTTTTTGAAACACAGTCT

ref|AC_000066.1| AGCAGTATTTTTATTGTACACACCTGGTGGGATATATGTGTTTTTTTTGAAACACAGTCT

************************************************************

ref|NC_000023.9| TGCTCTGTCACCCA

ref|AC_000066.1| TGCTCTGTCACCCA

**************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS8087(SHGC-152185)	NC_000023	152547714--152547748	17.5	100	70
	AC_000066	153128203--153128233	15.5	100	62
	AC_000155	141551962--141551995	17.0	100	68

DXS8087(SHGC-152185)	Information
Amplicon	130-132 bp
Forward ampliprimer	CTGCGCCAGTGAACAAGG
Reverse ampliprimer	GGGTCATGCACAAATGCAC

>ref|NC_000023.9|

CTGCGCCAGTGAACAAGGCAGGCCCATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTGTCTGTCTGTCTGTCTGCCTGTGCATG

TCTGTGTCTGTGTACACACATAAGTGCATTTGTGCATGACCC

>ref|AC_000066.1|

CTGCGCCAGTGAACAAGGCAGGCCCATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTGTCTGTCTGTCTGTCTGTCTGCCTGTGCATG

TCTGTGTCTGTGTACACACATAAGTGCATTTGTGCATGACCC

>ref|AC_000155.1|

CTGCGCCAGTGAACAAGGCAGGCCCGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTGTCTGTCTGTCTGTCTGCCTGTGCATGTC

TGTGTCTGTGTACACACATAAGTGCATTTGTGCATGACCC

DXS8087(SHGC-152185)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 132 bp

Sequence 2: ref|AC_000066.1| 132 bp

Sequence 3: ref|AC_000155.1| 130 bp

ref|NC_000023.9| CTGCGCCAGTGAACAAGGCAGGCCCATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|AC_000066.1| CTGCGCCAGTGAACAAGGCAGGCCCATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTG

ref|AC_000155.1| CTGCGCCAGTGAACAAGGCAGGCCC--GTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

************************* ****************************** **

ref|NC_000023.9| TCTGTCTGTCTGTCTGTCTGCCTGTGCATGTCTGTGTCTGTGTACACACATAAGTGCATT

ref|AC_000066.1| TCTGTCTGTCTGTCTGTCTGCCTGTGCATGTCTGTGTCTGTGTACACACATAAGTGCATT

ref|AC_000155.1| TCTGTCTGTCTGTCTGTCTGCCTGTGCATGTCTGTGTCTGTGTACACACATAAGTGCATT

************************************************************

ref|NC_000023.9| TGTGCATGACCC

ref|AC_000066.1| TGTGCATGACCC

ref|AC_000155.1| TGTGCATGACCC

************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152581.8	NC_000023	152581889--152581940	10.6	93	79
	AC_000066	153162379--153162430	10.6	93	79
	AC_000155	141586056--141586107	10.6	93	79

*WARNINGS!*
1.	The published HEMA152581.8 forward primer sequence (GGCAGGAGGATTACTTGAC) is missing a G residue at the 3’ end ^[1]. The correct sequence is GGCAGGAGGATTACTTGAGC. This primer pair matches two distinct locations on contig(s) from NCBI36/hg18 reference genome sequence by e-PCR: chrX:152581821+152581980 and chr15:24833223-24834846, but only one location from GRChH37/hg19 reference sequence: chrX:152928627+152928786 (Updated March 6, 2010).
2.	The locus HEMA152581.8 is, however, unique (UniSTS) and it can be developed into a marker using the, otherwise, amended primer pair shown below (Updated March 6, 2010).
^[1]	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.

HEMA152581.8	Information
Amplicon	132 bp
Forward ampliprimer	GGATTACTTGAGCCCAGGAG
Reverse ampliprimer	CCACCATGCTCGGTTCTAAT

>chrX:152581821+152581980 160bp GGCAGGAGGATTACTTGAGC AGCTGGGACTACAGGAGCAG

GGCAGGAGGATTACTTGAGCccaggagttcaacacccacgtgtgcagcat

agtgagactccatctctaaaaataaaataaaataaaataaaataaaataa

ataaaataaagtaaagtaaattagaaccgagcatggtggcCTGCTCCTGT

AGTCCCAGCT

>chr15:24833223-24834846 1624bp GGCAGGAGGATTACTTGAGC AGCTGGGACTACAGGAGCAG

GctgGGAGGATTACTTGAGCccaggagttcgaggctgcagtaagcccaat

tgtgccaccacactccggcctgggcaacagagcaacaccctgtctcaaaa

aaaagaaaaaaaaaacaaacgagatcatgtcctttacaacaacatagagt

tggagaccattatcctaagtgaactaatgcaggaacagaaaaccaaatac

tgcatgttctcacttacaagtgaggagccaaacactgagtacacacagac

aaatagaaaggaacaacagacaccaggcctatttgaaagtagagatgaaa

ggaaggtgagaatcaaaaaactacctatcggttactatgcttattacctg

ggtgatgaaataatctggacacaccaaatccccatgacatgaaatttacc

tatatgacaaacctacatctgtacccctgaagctaaaataaaagttggaa

gagggccgggcacagtggctcacgcctgtaatcccagcactttgggaggc

caaggtgggcagatcacttgagcccaagagtttgagaccagcctggcaac

atggtgagaccccatctctgctaaaaatacaaaaattagccaggcatggt

ggcgggcacctgtagtcccagctactcgggaggctgagtcacgagaattg

cttgaacccaggaggcagaggttgcaatgagcctagatagcgccactcca

ctccaacccagacgacaaagcgagaccctgtctcaaaaacaaacaaacaa

aaaaagttggaagacaaaaaaaaaactaggaatgtaaaaaaaagttttat

tgaagaaactttttccagattttgctttgagaaaatatatgtctagataa

agtccacaaacactggtccagatgagagtcaataactatatacaagatca

aatgatgttattctcatttaccctttctcttattcattacattcaaagct

ttgcctagaactggtaaaaggctggtaagattttgagatggtcactctga

tgcactgctggtgaacatatatatcaagttaatttttatggaaagtttgc

atcctttggacaagtagttccacttccaaaggtacaaattagggaaaaac

tgtcatggtaaattgaaattggccacaaactctaccaccaagggtagact

ctacttgatctaggtcagacttgcagcttgctttgactaacagaatatca

taaaagtgatgctgtatgaattctgaacaagacttgcttctagtctcatc

caaattccgactgctcctatatgaataagctcacactagacaccttgaga

cagagcaagcccctgtctcaaaattaaaagagtgaaattctgtcctttgc

agcaacgtggctagagctgaagcttcttccagttctagatcagaggcttg

agcacaacgaataagagaaagggtaaaagaggataacatgattttgtttc

ctatatagtaattgactcaaaagtttgttcaaggtctcacaatttctttt

taacttaaaaataaaaataatatctaaagaaaaaaatgaaatgctgcact

caaagcacactgcttcattaagatatttaacaatttaaagctgggcgtgg

tggcCTGCTCCTGTAGTCCCAGCT

>chrX:152581828+152581959 132bp GGATTACTTGAGCCCAGGAG CCACCATGCTCGGTTCTAAT

GGATTACTTGAGCCCAGGAGttcaacacccacgtgtgcagcatagtgaga

ctccatctctaaaaataaaataaaataaaataaaataaaataaataaaat

aaagtaaagtaaATTAGAACCGAGCATGGTGG

>ref|NC_000023.9|

GGATTACTTGAGCCCAGGAGTTCAACACCCACGTGTGCAGCATAGTGAGACTCCATCTCTAAAAATAAAATAAAATAAAATAAAATAAAA

TAAATAAAATAAAGTAAAGTAAATTAGAACCGAGCATGGTGG

>ref|AC_000066.1|

GGATTACTTGAGCCCAGGAGTTCAACACCCACGTGTGCAGCATAGTGAGACTCCATCTCTAAAAATAAAATAAAATAAAATAAAATAAAA

TAAATAAAATAAAGTAAAGTAAATTAGAACCGAGCATGGTGG

>ref|AC_000155.1|

GGATTACTTGAGCCCAGGAGTTCAACACCCACGTGTGCAGCATAGTGAGACTCCATCTCTAAAAATAAAATAAAATAAAATAAAATAAAA

TAAATAAAATAAAGTAAAGTAAATTAGAACCGAGCATGGTGG

HEMA152581.8

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 132 bp

Sequence 2: ref|AC_000066.1| 132 bp

Sequence 3: ref|AC_000155.1| 132 bp

ref|NC_000023.9| GGATTACTTGAGCCCAGGAGTTCAACACCCACGTGTGCAGCATAGTGAGACTCCATCTCT

ref|AC_000155.1| GGATTACTTGAGCCCAGGAGTTCAACACCCACGTGTGCAGCATAGTGAGACTCCATCTCT

ref|AC_000066.1| GGATTACTTGAGCCCAGGAGTTCAACACCCACGTGTGCAGCATAGTGAGACTCCATCTCT

************************************************************

ref|NC_000023.9| AAAAATAAAATAAAATAAAATAAAATAAAATAAATAAAATAAAGTAAAGTAAATTAGAAC

ref|AC_000155.1| AAAAATAAAATAAAATAAAATAAAATAAAATAAATAAAATAAAGTAAAGTAAATTAGAAC

ref|AC_000066.1| AAAAATAAAATAAAATAAAATAAAATAAAATAAATAAAATAAAGTAAAGTAAATTAGAAC

************************************************************

ref|NC_000023.9| CGAGCATGGTGG

ref|AC_000155.1| CGAGCATGGTGG

ref|AC_000066.1| CGAGCATGGTGG

************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**PNCKInt2**	NC_000023	152591604--152591641	18.0	89	58
	AC_000066	153172094--153172131	18.0	89	58
	AC_000155	141595771--141595808	18.0	89	58

PNCKInt2	Information
Amplicon	179 bp
Forward ampliprimer	CATCATCTCACACCCACTCG
Reverse ampliprimer	AGACATGCTGCTGCTGAAGA

>ref|NC_000023.9|

CATCATCTCACACCCACTCGCGTAGTCACACACACATGCCATCTCACACAGGTACACACACACACACACACACACACACACGATCACACA

CGCGCGCGCACACTCACGAGCCGAGCCTCTCGCGGATCTCGTAGACGCTGCTGATGTCCTCCGTGTGTTTCTTCAGCAGCAGCATGTCT

>ref|AC_000066.1|

CATCATCTCACACCCACTCGCGTAGTCACACACACATGCCATCTCACACAGGTACACACACACACACACACACACACACACGATCACACA

CGCGCGCGCACACTCACGAGCCGAGCCTCTCGCGGATCTCGTAGACGCTGCTGATGTCCTCCGTGTGTTTCTTCAGCAGCAGCATGTCT

>ref|AC_000155.1|

CATCATCTCACACCCACTCGCGTAGTCACACACACATGCCATCTCACACAGGTACACACACACACACACACACACACACACGATCACACA

CGCGCGCGCACACTCACGAGCCGAGCCTCTCGCGGATCTCGTAGACGCTGCTGATGTCCTCCGTGTGTTTCTTCAGCAGCAGCATGTCT

PNCKInt2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 179 bp

Sequence 2: ref|AC_000066.1| 179 bp

Sequence 3: ref|AC_000155.1| 179 bp

ref|NC_000023.9| CATCATCTCACACCCACTCGCGTAGTCACACACACATGCCATCTCACACAGGTACACACA

ref|AC_000155.1| CATCATCTCACACCCACTCGCGTAGTCACACACACATGCCATCTCACACAGGTACACACA

ref|AC_000066.1| CATCATCTCACACCCACTCGCGTAGTCACACACACATGCCATCTCACACAGGTACACACA

************************************************************

ref|NC_000023.9| CACACACACACACACACACACGATCACACACGCGCGCGCACACTCACGAGCCGAGCCTCT

ref|AC_000155.1| CACACACACACACACACACACGATCACACACGCGCGCGCACACTCACGAGCCGAGCCTCT

ref|AC_000066.1| CACACACACACACACACACACGATCACACACGCGCGCGCACACTCACGAGCCGAGCCTCT

************************************************************

ref|NC_000023.9| CGCGGATCTCGTAGACGCTGCTGATGTCCTCCGTGTGTTTCTTCAGCAGCAGCATGTCT

ref|AC_000155.1| CGCGGATCTCGTAGACGCTGCTGATGTCCTCCGTGTGTTTCTTCAGCAGCAGCATGTCT

ref|AC_000066.1| CGCGGATCTCGTAGACGCTGCTGATGTCCTCCGTGTGTTTCTTCAGCAGCAGCATGTCT

***********************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**SLC6A8Exon1**	NC_000023	152606681--152606713	11.0	93	57
	NC_000023	152606920--152606964	16.0	77	51
	AC_000066	153187171--153187203	11.0	93	57
	AC_000066	153187410--153187454	16.0	77	51

SLC6A8Exon1	Information
Amplicon	439 bp
Forward ampliprimer	GACGGGGAAGAGAGGGATAG
Reverse ampliprimer	AGCCTGAAGAATCTCATGTGTG

>ref|NC_000023.9|

GACGGGGAAGAGAGGGATAGTCGGAGCGAGGTGGCGAGTCGCTGAGCCCGCCGCGGCCCCGAGAGCGGCTGCAGCCGCCGCCGCCGGGAAGG

AGAGGGCGAGGCGCGCCCGAGCCGCCGCCGCCGCCGCCACCGCCGCCGCCGCCACCACCGCCACCGGAGTCGCGGGCCAGCCGGGCAGCCTC

CGCGGGCCCCGGCCGGGGCGGGGGGCGCGGGCCACAGGCCCCTGCTCCGGCCGCCGCTTGCAGACCGCGGGCGCCGATGTCGCCCGCGCCCC

GCTAGGCTGAGCCTCGGGTCGGGCGAGGAGCCGCCGCAGCCGCCGCCGCCCGAGCCGCGGGCAGGAGCCTCGGGAGCCGCCGCCGCCGCCGC

CGCCGCCCGGCCGGGCCCCGCCGCCGCCCGCGCGCCCCCGGGCCCCCGACACACATGAGATTCTTCAGGCT

>ref|AC_000066.1|

GACGGGGAAGAGAGGGATAGTCGGAGCGAGGTGGCGAGTCGCTGAGCCCGCCGCGGCCCCGAGAGCGGCTGCAGCCGCCGCCGCCGGGAAGG

AGAGGGCGAGGCGCGCCCGAGCCGCCGCCGCCGCCGCCACCGCCGCCGCCGCCACCACCGCCACCGGAGTCGCGGGCCAGCCGGGCAGCCTC

CGCGGGCCCCGGCCGGGGCGGGGGGCGCGGGCCACAGGCCCCTGCTCCGGCCGCCGCTTGCAGACCGCGGGCGCCGATGTCGCCCGCGCCCC

GCTAGGCTGAGCCTCGGGTCGGGCGAGGAGCCGCCGCAGCCGCCGCCGCCCGAGCCGCGGGCAGGAGCCTCGGGAGCCGCCGCCGCCGCCGC

CGCCGCCCGGCCGGGCCCCGCCGCCGCCCGCGCGCCCCCGGGCCCCCGACACACATGAGATTCTTCAGGCT

SLC6A8Exon1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 439 bp

Sequence 2: ref|AC_000066.1| 439 bp

ref|NC_000023.9| GACGGGGAAGAGAGGGATAGTCGGAGCGAGGTGGCGAGTCGCTGAGCCCGCCGCGGCCCC

ref|AC_000066.1| GACGGGGAAGAGAGGGATAGTCGGAGCGAGGTGGCGAGTCGCTGAGCCCGCCGCGGCCCC

************************************************************

ref|NC_000023.9| GAGAGCGGCTGCAGCCGCCGCCGCCGGGAAGGAGAGGGCGAGGCGCGCCCGAGCCGCCGC

ref|AC_000066.1| GAGAGCGGCTGCAGCCGCCGCCGCCGGGAAGGAGAGGGCGAGGCGCGCCCGAGCCGCCGC

************************************************************

ref|NC_000023.9| CGCCGCCGCCACCGCCGCCGCCGCCACCACCGCCACCGGAGTCGCGGGCCAGCCGGGCAG

ref|AC_000066.1| CGCCGCCGCCACCGCCGCCGCCGCCACCACCGCCACCGGAGTCGCGGGCCAGCCGGGCAG

************************************************************

ref|NC_000023.9| CCTCCGCGGGCCCCGGCCGGGGCGGGGGGCGCGGGCCACAGGCCCCTGCTCCGGCCGCCG

ref|AC_000066.1| CCTCCGCGGGCCCCGGCCGGGGCGGGGGGCGCGGGCCACAGGCCCCTGCTCCGGCCGCCG

************************************************************

ref|NC_000023.9| CTTGCAGACCGCGGGCGCCGATGTCGCCCGCGCCCCGCTAGGCTGAGCCTCGGGTCGGGC

ref|AC_000066.1| CTTGCAGACCGCGGGCGCCGATGTCGCCCGCGCCCCGCTAGGCTGAGCCTCGGGTCGGGC

************************************************************

ref|NC_000023.9| GAGGAGCCGCCGCAGCCGCCGCCGCCCGAGCCGCGGGCAGGAGCCTCGGGAGCCGCCGCC

ref|AC_000066.1| GAGGAGCCGCCGCAGCCGCCGCCGCCCGAGCCGCGGGCAGGAGCCTCGGGAGCCGCCGCC

************************************************************

ref|NC_000023.9| GCCGCCGCCGCCGCCCGGCCGGGCCCCGCCGCCGCCCGCGCGCCCCCGGGCCCCCGACAC

ref|AC_000066.1| GCCGCCGCCGCCGCCCGGCCGGGCCCCGCCGCCGCCCGCGCGCCCCCGGGCCCCCGACAC

************************************************************

ref|NC_000023.9| ACATGAGATTCTTCAGGCT

ref|AC_000066.1| ACATGAGATTCTTCAGGCT

*******************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**PDZD4Int1**	NC_000023	152741273--152741325	26.5	84	70
	AC_000066	153321757--153321811	27.5	84	74
	AC_000155	141745081--141745135	27.5	84	74

PDZD4Int1	Information
Amplicon	235-237 bp
Forward ampliprimer	CCAACTCCTGTGCAAAATGA
Reverse ampliprimer	GAGTGGGAGGATTGCTTGAA

>ref|NC_000023.9|

CCAACTCCTGTGCAAAATGACCACATATGCCTCCGCTTTGTTCCCTGTCCATCTGTTTTTTCTTTCTCTCTCTCTCTCTCTCTCCCTCCC

TCCCTCTCTCTCTCTCTCTTTCTCTCTGTCTTTCTTTCTTTTTTTTCCAGACACAGTCTGGCTCCATTGCCCAGGCTAGAGTGCAGTGGT

ACAATCACGGCTCACAGAAGCCTCCAACTCCTGGGTTCAAGCAATCCTCCCACTC

>ref|AC_000066.1|

CCAACTCCTGTGCAAAATGACCACATATGCCTCCGCTTTGTTCCCTGTCCATCTGTTTTTTCTTTCTCTCTCTCTCTCTCTCTCCCTCCC

TCCCTCTCTCTCTCTCTCTCTTTCTCTCTGTCTTTCTTTCTTTTTTTTCCAGACACAGTCTGGCTCCATTGCCCAGGCTAGAGTGCAGTG

GTACAATCACGGCTCACAGAAGCCTCCAACTCCTGGGTTCAAGCAATCCTCCCACTC

>ref|AC_000155.1|

CCAACTCCTGTGCAAAATGACCACATATGCCTCCGCTTTGTTCCCTGTCCATCTGTTTTTTCTTTCTCTCTCTCTCTCTCTCTCCCTCCC

TCCCTCTCTCTCTCTCTCTCTTTCTCTCTGTCTTTCTTTCTTTTTTTTCCAGACACAGTCTGGCTCCATTGCCCAGGCTAGAGTGCAGTG

GTACAATCACGGCTCACAGAAGCCTCCAACTCCTGGGTTCAAGCAATCCTCCCACTC

PDZD4Int1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 235 bp

Sequence 2: ref|AC_000066.1| 237 bp

Sequence 3: ref|AC_000155.1| 237 bp

ref|AC_000066.1| CCAACTCCTGTGCAAAATGACCACATATGCCTCCGCTTTGTTCCCTGTCCATCTGTTTTT

ref|AC_000155.1| CCAACTCCTGTGCAAAATGACCACATATGCCTCCGCTTTGTTCCCTGTCCATCTGTTTTT

ref|NC_000023.9| CCAACTCCTGTGCAAAATGACCACATATGCCTCCGCTTTGTTCCCTGTCCATCTGTTTTT

************************************************************

ref|AC_000066.1| TCTTTCTCTCTCTCTCTCTCTCTCCCTCCCTCCCTCTCTCTCTCTCTCTCTTTCTCTCTG

ref|AC_000155.1| TCTTTCTCTCTCTCTCTCTCTCTCCCTCCCTCCCTCTCTCTCTCTCTCTCTTTCTCTCTG

ref|NC_000023.9| TCTTTCTCTCTCTCTCTCTCTCTCCCTCCCTCCCTCTCTCTCTCTCTC--TTTCTCTCTG

************************************************ **********

ref|AC_000066.1| TCTTTCTTTCTTTTTTTTCCAGACACAGTCTGGCTCCATTGCCCAGGCTAGAGTGCAGTG

ref|AC_000155.1| TCTTTCTTTCTTTTTTTTCCAGACACAGTCTGGCTCCATTGCCCAGGCTAGAGTGCAGTG

ref|NC_000023.9| TCTTTCTTTCTTTTTTTTCCAGACACAGTCTGGCTCCATTGCCCAGGCTAGAGTGCAGTG

************************************************************

ref|AC_000066.1| GTACAATCACGGCTCACAGAAGCCTCCAACTCCTGGGTTCAAGCAATCCTCCCACTC

ref|AC_000155.1| GTACAATCACGGCTCACAGAAGCCTCCAACTCCTGGGTTCAAGCAATCCTCCCACTC

ref|NC_000023.9| GTACAATCACGGCTCACAGAAGCCTCCAACTCCTGGGTTCAAGCAATCCTCCCACTC

*********************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN87871	NC_000023	152769114--152769164	12.8	100	102
	AC_000066	153349600--153349650	12.8	100	102
	AC_000155	141772462--141772500	9.8	100	78

*WARNING!*
To improve specificity (i.e., reducing the chances of amplification of head-to-head primers annealing to more than one location on contig(s) from NCBI36/hg18 reference genome sequence by e-PCR, the published ^[1] REN87871 primer pair sequences (CTCAACTCACAGCAGCCTCA and CTCTTGAGCCCAGGAGTTTG) are amended to the sequences shown below. The locus REN87871 can thus be developed into a UniSTS marker (Updated March 6, 2010).
^[1]	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.

REN87871	Information
Amplicon	101-120 bp
Forward ampliprimer	TCGCCTCGCTAATTTTTCTT
Reverse ampliprimer	CTGGGCAACATAGCAAGACC

>chrX:152769100+152769219 120bp TCGCCTCGCTAATTTTTCTT CTGGGCAACATAGCAAGACC

TCGCCTCGCTAATTTTTCTTtctttctttctttctttctttctttctttc

tttctttctttctttttttttttttttttttttgctttttgtagagacag

GGTCTTGCTATGTTGCCCAG

>ref|NC_000023.9|

TCGCCTCGCTAATTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTTTTTTTTTTTTTGCTTTTT

GTAGAGACAGGGTCTTGCTATGTTGCCCAG

>ref|AC_000066.1|

TCGCCTCGCTAATTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTTTTTTTTTTTTTGCTTTTT

GTAGAGACAGGGTCTTGCTATGTTGCCCAG

>ref|AC_000155.1|

TCGCCTCGCTAATTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTTTTTTGCTTTTTGTAGAGACAGGGTCTTGCT

ATGTTGCCCAG

REN87871

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 120 bp

Sequence 2: ref|AC_000066.1| 120 bp

Sequence 3: ref|AC_000155.1| 101 bp

ref|NC_000023.9| TCGCCTCGCTAATTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

ref|AC_000066.1| TCGCCTCGCTAATTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

ref|AC_000155.1| TCGCCTCGCTAATTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTT-------

*****************************************************

ref|NC_000023.9| TCTTTTTTTTTTTTTTTTTTTTTGCTTTTTGTAGAGACAGGGTCTTGCTATGTTGCCCAG

ref|AC_000066.1| TCTTTTTTTTTTTTTTTTTTTTTGCTTTTTGTAGAGACAGGGTCTTGCTATGTTGCCCAG

ref|AC_000155.1| ------------TTTTTTTTTTTGCTTTTTGTAGAGACAGGGTCTTGCTATGTTGCCCAG

************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152778.1	NC_000023	152778106--152778183	19.0	92	129
	AC_000066	153358592--153358669	19.0	92	129
	AC_000155	141781434--141781511	19.0	92	129

HEMA152778.1	Information
Amplicon	232 bp
Forward ampliprimer	CTCTAGCCTGGGTGATCGAG
Reverse ampliprimer	TCAGCTGAAGACCCATTTCC

>ref|NC_000023.9|

CTCTAGCCTGGGTGATCGAGCGAGACTCCATCTCAAAAAAGAAAAAAAGAAAGAAAGAAAGAAAGGAAGGAAGGAAGGAAGGAAGGAAGG

AAGGAAGGAAGGGAAGGAAGGAAGGAAGGGAAGGAAGGAAGGAAGAAAGGCGCCAGCACAGGTTTGCCCTGCAGGTATGGGTGCACTCAT

GCCCCACCACCCTACTGGGTGCATAGAGGCTGGGAAATGGGTCTTCAGCTGA

>ref|AC_000066.1|

CTCTAGCCTGGGTGATCGAGCGAGACTCCATCTCAAAAAAGAAAAAAAGAAAGAAAGAAAGAAAGGAAGGAAGGAAGGAAGGAAGGAAGG

AAGGAAGGAAGGGAAGGAAGGAAGGAAGGGAAGGAAGGAAGGAAGAAAGGCGCCAGCACAGGTTTGCCCTGCAGGTATGGGTGCACTCAT

GCCCCACCACCCTACTGGGTGCATAGAGGCTGGGAAATGGGTCTTCAGCTGA

>ref|AC_000155.1|

CTCTAGCCTGGGTGATCGAGCGAGACTCCATCTCAAAAAAGAAAAAAAGAAAGAAAGAAAGAAAGGAAGGAAGGAAGGAAGGAAGGAAGG

AAGGAAGGAAGGGAAGGAAGGAAGGAAGGGAAGGAAGGAAGGAAGAAAGGCGCCAGCACAGGTTTGCCCTGCAGGTATGGGTGCACTCAT

GCCCCACCACCCTACTGGGTGCATAGAGGCTGGGAAATGGGTCTTCAGCTGA

HEMA152778.1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 232 bp

Sequence 2: ref|AC_000066.1| 232 bp

Sequence 3: ref|AC_000155.1| 232 bp

ref|NC_000023.9| CTCTAGCCTGGGTGATCGAGCGAGACTCCATCTCAAAAAAGAAAAAAAGAAAGAAAGAAA

ref|AC_000155.1| CTCTAGCCTGGGTGATCGAGCGAGACTCCATCTCAAAAAAGAAAAAAAGAAAGAAAGAAA

ref|AC_000066.1| CTCTAGCCTGGGTGATCGAGCGAGACTCCATCTCAAAAAAGAAAAAAAGAAAGAAAGAAA

************************************************************

ref|NC_000023.9| GAAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGGAAGGAAGGAAGGAAGGG

ref|AC_000155.1| GAAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGGAAGGAAGGAAGGAAGGG

ref|AC_000066.1| GAAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGAAGGGAAGGAAGGAAGGAAGGG

************************************************************

ref|NC_000023.9| AAGGAAGGAAGGAAGAAAGGCGCCAGCACAGGTTTGCCCTGCAGGTATGGGTGCACTCAT

ref|AC_000155.1| AAGGAAGGAAGGAAGAAAGGCGCCAGCACAGGTTTGCCCTGCAGGTATGGGTGCACTCAT

ref|AC_000066.1| AAGGAAGGAAGGAAGAAAGGCGCCAGCACAGGTTTGCCCTGCAGGTATGGGTGCACTCAT

************************************************************

ref|NC_000023.9| GCCCCACCACCCTACTGGGTGCATAGAGGCTGGGAAATGGGTCTTCAGCTGA

ref|AC_000155.1| GCCCCACCACCCTACTGGGTGCATAGAGGCTGGGAAATGGGTCTTCAGCTGA

ref|AC_000066.1| GCCCCACCACCCTACTGGGTGCATAGAGGCTGGGAAATGGGTCTTCAGCTGA

****************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN87914	NC_000023	152779117--152779297	45.3	83	185
	NC_000023	152779316--152779411	24.0	91	102
	AC_000066	153359603--153359787	46.3	83	193
	AC_000066	153359806--153359901	24.0	91	102
	AC_000155	141782445--141782621	44.3	82	177
	AC_000155	141782640--141782733	23.5	91	98

REN87914	Information
Amplicon	350-360 bp
Forward ampliprimer	AAGTGGAGGGCTCACCTGT
Reverse ampliprimer	GCGAGACAAAGAAAGAAAAAGAA

>ref|NC_000023.9|

AAGTGGAGGGCTCACCTGTGCTGGGTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTT

TTCTTTCTTTCTTTCTCTTTCTTTCCTTTTCTTTCTCTCCCTCCCTCCCTTCCTTTCTTTCTTTCTTGCTTGCTTTCTTGCTTTGTCTTT

CTCTTTCTTTCTTCTTTCTTTCTTTCTCTTTCCCTCCCTCCCTCCCTCTCTCTCTCTGTCTCTCTCTCTCTTTCTTTCTTTCTCTCTTTC

CCTCCCTCCCTCCCTCCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTTTCTTTCTTTCTTTTTCTTTCTTTGTCTCGC

>ref|AC_000066.1|

AAGTGGAGGGCTCACCTGTGCTGGGTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTC

TTTTTTCTTTCTTTCTTTCTCTTTCTTTCCTTTTCTTTCTCTCCCTCCCTCCCTTCCTTTCTTTCTTTCTTGCTTGCTTTCTTGCTTTGT

CTTTCTCTTTCTTTCTTCTTTCTTTCTTTCTCTTTCCCTCCCTCCCTCCCTCTCTCTCTCTGTCTCTCTCTCTCTTTCTTTCTTTCTCTC

TTTCCCTCCCTCCCTCCCTCCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTTTCTTTCTTTCTTTTTCTTTCTTTGTCTCGC

>ref|AC_000155.1|

AAGTGGAGGGCTCACCTGTGCTGGGTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTCT

TTCTTTCTTTCTCTTTCTTTCCTTTTCTTTCTCTCCCTCCCTCCCTTCCTTTCTTTCTTTCTTGCTTGCTTTCTTGCTTTGTCTTTCTCT

TTCTTTCTTCTTTCTTTCTTTCTCTTTCCCTCCCTCCCTCCCTCTCTCTCTCTGTCTCTCTCTCTCTTTCTTTCTTTCTCTCTTTCCCTC

CCTCCCTCCCTCCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTTTCTTTCTTTCTTTTTCTTTCTTTGTCTCGC

REN87914

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 356 bp

Sequence 2: ref|AC_000066.1| 360 bp

Sequence 3: ref|AC_000155.1| 350 bp

ref|AC_000066.1| AAGTGGAGGGCTCACCTGTGCTGGGTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

ref|NC_000023.9| AAGTGGAGGGCTCACCTGTGCTGGGTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

ref|AC_000155.1| AAGTGGAGGGCTCACCTGTGCTGGGTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

************************************************************

ref|AC_000066.1| TCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTCTTTCTTTCTTTCTCTTTCTTTCC

ref|NC_000023.9| TCTTTCTTTCTTTCTTTCTTTCTTTC----TTTTTTCTTTCTTTCTTTCTCTTTCTTTCC

ref|AC_000155.1| TCTTTCTTTCTTTCTTTCTTTC--------TTTTTTCTTTCTTTCTTTCTCTTTCTTTCC

********************** ******************************

ref|AC_000066.1| TTTTCTTTCTCTCCCTCCCTCCCTTCCTTTCTTTCTTTCTTGCTTGCTTTCTTGCTTTGT

ref|NC_000023.9| TTTTCTTTCTCTCCCTCCCTCCCTTCCTTTCTTTCTTTCTTGCTTGCTTTCTTGCTTTGT

ref|AC_000155.1| TTTTCTTTCTCTCCCTCCCTCCCTTCCTTTCTTTCTTTCTTGCTTGCTTTCTTGCTTTGT

************************************************************

ref|AC_000066.1| CTTTCTCTTTCTTTCTTCTTTCTTTCTTTCTCTTTCCCTCCCTCCCTCCCTCTCTCTCTC

ref|NC_000023.9| CTTTCTCTTTCTTTCTTCTTTCTTTCTTTCTCTTTCCCTCCCTCCCTCCCTCTCTCTCTC

ref|AC_000155.1| CTTTCTCTTTCTTTCTTCTTTCTTTCTTTCTCTTTCCCTCCCTCCCTCCCTCTCTCTCTC

************************************************************

ref|AC_000066.1| TGTCTCTCTCTCTCTTTCTTTCTTTCTCTCTTTCCCTCCCTCCCTCCCTCCCTCTCTCTC

ref|NC_000023.9| TGTCTCTCTCTCTCTTTCTTTCTTTCTCTCTTTCCCTCCCTCCCTCCCTCCCTCTCTCTC

ref|AC_000155.1| TGTCTCTCTCTCTCTTTCTTTCTTTCTCTCTTTCCCTCCCTCCCTCCCTCCCTCTCTCTC

************************************************************

ref|AC_000066.1| TCTCTCTCTCTCTCTCTCTCTCTCTTTCTTTCTTTCTTTCTTTTTCTTTCTTTGTCTCGC

ref|NC_000023.9| TCTCTCTCTCTCTCTCTCTCTCTCTTTCTTTCTTTCTTTCTTTTTCTTTCTTTGTCTCGC

ref|AC_000155.1| TCTCTCTCTCTCTCTCTCTCTCT--TTCTTTCTTTCTTTCTTTTTCTTTCTTTGTCTCGC

*********************** ***********************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS9796(REN88038)	NC_000023	152807881--152807905	12.5	91	41
	NC_000023	152807905--152807923	9.5	100	38
	AC_000066	153388377--153388401	12.5	91	41
	AC_000066	153388401--153388419	9.5	100	38
	AC_000155	141811322--141811346	12.5	91	41
	AC_000155	141811346--141811364	9.5	100	38

DXS9796(REN88038)	Information
Amplicon	153 bp
Forward ampliprimer	CGCAGCATACTTTTTGAGCTT
Reverse ampliprimer	ACAAACCCTGATCCAACCAA

>ref|NC_000023.9|

CGCAGCATACTTTTTGAGCTTCATCCATGTTGGTGTGTGTGTTTGTGTGTGTGTGTGAGAGAGAGAGAGAGAGAGCATGCGGGCGAGAGA

GAGCAGTATATTTCCTTTTGATTGCCGAGTAGTATTCCATTGTTTGGTTGGATCAGGGTTTGT

>ref|AC_000066.1|

CGCAGCATACTTTTTGAGCTTCATCCATGTTGGTGTGTGTGTTTGTGTGTGTGTGTGAGAGAGAGAGAGAGAGAGCATGCGGGCGAGAGA

GAGCAGTATATTTCCTTTTGATTGCCGAGTAGTATTCCATTGTTTGGTTGGATCAGGGTTTGT

>ref|AC_000155.1|

CGCAGCATACTTTTTGAGCTTCATCCATGTTGGTGTGTGTGTTTGTGTGTGTGTGTGAGAGAGAGAGAGAGAGAGCATGCGGGCGAGAGA

GAGCAGTATATTTCCTTTTGATTGCCGAGTAGTATTCCATTGTTTGGTTGGATCAGGGTTTGT

DXS9796(REN88038)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 153 bp

Sequence 2: ref|AC_000066.1| 153 bp

Sequence 3: ref|AC_000155.1| 153 bp

ref|AC_000066.1| CGCAGCATACTTTTTGAGCTTCATCCATGTTGGTGTGTGTGTTTGTGTGTGTGTGTGAGA

ref|NC_000023.9| CGCAGCATACTTTTTGAGCTTCATCCATGTTGGTGTGTGTGTTTGTGTGTGTGTGTGAGA

ref|AC_000155.1| CGCAGCATACTTTTTGAGCTTCATCCATGTTGGTGTGTGTGTTTGTGTGTGTGTGTGAGA

************************************************************

ref|AC_000066.1| GAGAGAGAGAGAGAGCATGCGGGCGAGAGAGAGCAGTATATTTCCTTTTGATTGCCGAGT

ref|NC_000023.9| GAGAGAGAGAGAGAGCATGCGGGCGAGAGAGAGCAGTATATTTCCTTTTGATTGCCGAGT

ref|AC_000155.1| GAGAGAGAGAGAGAGCATGCGGGCGAGAGAGAGCAGTATATTTCCTTTTGATTGCCGAGT

************************************************************

ref|AC_000066.1| AGTATTCCATTGTTTGGTTGGATCAGGGTTTGT

ref|NC_000023.9| AGTATTCCATTGTTTGGTTGGATCAGGGTTTGT

ref|AC_000155.1| AGTATTCCATTGTTTGGTTGGATCAGGGTTTGT

*********************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88052	NC_000023	152810851--152810880	15.0	100	60
	AC_000066	153391347--153391374	14.0	100	56
	AC_000155	141814292--141814321	15.0	100	60

REN88052	Information
Amplicon	100-102 bp
Forward ampliprimer	TGCTGCTTCCTCCTTGTTTT
Reverse ampliprimer	GCTGTTGAGTGTCCTCACGA

>ref|NC_000023.9|

TGCTGCTTCCTCCTTGTTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCGGATTGCTCTGTTGGGTGAAGCCAGGTGCCACATCGTGAGG

ACACTCAACAGC

>ref|AC_000066.1|

TGCTGCTTCCTCCTTGTTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCGGATTGCTCTGTTGGGTGAAGCCAGGTGCCACATCGTGAGGAC

ACTCAACAGC

>ref|AC_000155.1|

TGCTGCTTCCTCCTTGTTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCGGATTGCTCTGTTGGGTGAAGCCAGGTGCCACATCGTGAGG

ACACTCAACAGC

REN88052

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 102 bp

Sequence 2: ref|AC_000066.1| 100 bp

Sequence 3: ref|AC_000155.1| 102 bp

ref|NC_000023.9| TGCTGCTTCCTCCTTGTTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCGGATTGCTCTG

ref|AC_000155.1| TGCTGCTTCCTCCTTGTTTTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCGGATTGCTCTG

ref|AC_000066.1| TGCTGCTTCCTCCTTGTTTTCTCTCTCTCTCTCTCTCTCTCTCTCTC--GGATTGCTCTG

*********************************************** ***********

ref|NC_000023.9| TTGGGTGAAGCCAGGTGCCACATCGTGAGGACACTCAACAGC

ref|AC_000155.1| TTGGGTGAAGCCAGGTGCCACATCGTGAGGACACTCAACAGC

ref|AC_000066.1| TTGGGTGAAGCCAGGTGCCACATCGTGAGGACACTCAACAGC

******************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88058	NC_000023	152812156--152812299	36.8	82	176
	AC_000066	153392650--153392785	34.8	80	160
	AC_000155	141815597--141815732	34.8	80	160

REN88058	Information
Amplicon	279-287 bp
Forward ampliprimer	TCCAACCTGGGTGACAGAGT
Reverse ampliprimer	TTCTGGTGAGGGCTGTCTTC

>ref|NC_000023.9|

TCCAACCTGGGTGACAGAGTAAGACCCTGCCTCAAAAGAAAAGAAAGAAAGAGAGAGAAGGAAAGAAGAAAGAAAGAAAGAAAGAAAGAA

AGAAAGAAAGAAAGAAAGAGAAAGAAGGAAAGAAAGAAAGAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAAAGAAAGAGAGAAAGAAAAA

GAACACAAGAAAAGCACTGTTCCCTCCCCTCCACCACTGCCCCTACCATGTGAGGACACAGTGAGAAGGTGGCTGTCTCTAAACCAGGAA

GACAGCCCTCACCAGAA

>ref|AC_000066.1|

TCCAACCTGGGTGACAGAGTAAGACCCTGCCTCAAAAGAAAAGAAAGAAAGAGAGAGAAGGAAAGAAGAAAGAAAGAAAGAAAGAAAGAA

AGAAAGAAAGAGAAAGAAGGAAAGAAAGAAAGAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAAAGAAAGAGAGAAAGAAAAAGAACACAA

GAAAAGCACTGTTCCCTCCCCTCCACCACTGCCCCTACCATGTGAGGACACAGTGAGAAGGTGGCTGTCTCTAAACCAGGAAGACAGCCC

TCACCAGAA

>ref|AC_000155.1|

TCCAACCTGGGTGACAGAGTAAGACCCTGCCTCAAAAGAAAAGAAAGAAAGAGAGAGAAGGAAAGAAGAAAGAAAGAAAGAAAGAAAGAA

AGAAAGAAAGAGAAAGAAGGAAAGAAAGAAAGAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAAAGAAAGAGAGAAAGAAAAAGAACACAA

GAAAAGCACTGTTCCCTCCCCTCCACCACTGCCCCTACCATGTGAGGACACAGTGAGAAGGTGGCTGTCTCTAAACCAGGAAGACAGCCC

TCACCAGAA

REN88058

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 287 bp

Sequence 2: ref|AC_000066.1| 279 bp

Sequence 3: ref|AC_000155.1| 279 bp

ref|AC_000066.1| TCCAACCTGGGTGACAGAGTAAGACCCTGCCTCAAAAGAAAAGAAAGAAAGAGAGAGAAG

ref|AC_000155.1| TCCAACCTGGGTGACAGAGTAAGACCCTGCCTCAAAAGAAAAGAAAGAAAGAGAGAGAAG

ref|NC_000023.9| TCCAACCTGGGTGACAGAGTAAGACCCTGCCTCAAAAGAAAAGAAAGAAAGAGAGAGAAG

************************************************************

ref|AC_000066.1| GAAAGAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA--------GAAAGAAGGAA

ref|AC_000155.1| GAAAGAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGA--------GAAAGAAGGAA

ref|NC_000023.9| GAAAGAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAGGAA

***************************************** ***********

ref|AC_000066.1| AGAAAGAAAGAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAAAGAAAGAGAGAAAGAAAAA

ref|AC_000155.1| AGAAAGAAAGAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAAAGAAAGAGAGAAAGAAAAA

ref|NC_000023.9| AGAAAGAAAGAGAAAGAAAGAAAGAAAGAAAGAGAAAGAAAAGAAAGAGAGAAAGAAAAA

************************************************************

ref|AC_000066.1| GAACACAAGAAAAGCACTGTTCCCTCCCCTCCACCACTGCCCCTACCATGTGAGGACACA

ref|AC_000155.1| GAACACAAGAAAAGCACTGTTCCCTCCCCTCCACCACTGCCCCTACCATGTGAGGACACA

ref|NC_000023.9| GAACACAAGAAAAGCACTGTTCCCTCCCCTCCACCACTGCCCCTACCATGTGAGGACACA

************************************************************

ref|AC_000066.1| GTGAGAAGGTGGCTGTCTCTAAACCAGGAAGACAGCCCTCACCAGAA

ref|AC_000155.1| GTGAGAAGGTGGCTGTCTCTAAACCAGGAAGACAGCCCTCACCAGAA

ref|NC_000023.9| GTGAGAAGGTGGCTGTCTCTAAACCAGGAAGACAGCCCTCACCAGAA

***********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
RENBPInt9.1	NC_000023	152855845--152855891	11.8	100	94
	AC_000066	153436368--153436414	11.8	100	94
	AC_000155	141855356--141855402	11.8	100	94

RENBPInt9.1	Information
Amplicon	193 bp
Forward ampliprimer	CTGCACTGAGCCATGATTGT
Reverse ampliprimer	CCTGTGTCATGTCAAGCAGTC

>ref|NC_000023.9|

CTGCACTGAGCCATGATTGTGCCACTGCACTCAGCCTGGGCGATAGAGACCCTGTCTCAAAAATAAATAAATAAATAAATAAATAAATAA

ATAAATAAATAAATAAAAACTAGTTCTGTGTGTATTCATTCTGTCTTTTGAGTTACATTGTTCAAGTCCTTACTTATTTTTTGACTGCTT

GACATGACACAGG

>ref|AC_000066.1|

CTGCACTGAGCCATGATTGTGCCACTGCACTCAGCCTGGGCGATAGAGACCCTGTCTCAAAAATAAATAAATAAATAAATAAATAAATAA

ATAAATAAATAAATAAAAACTAGTTCTGTGTGTATTCATTCTGTCTTTTGAGTTACATTGTTCAAGTCCTTACTTATTTTTTGACTGCTT

GACATGACACAGG

>ref|AC_000155.1|

CTGCACTGAGCCATGATTGTGCCACTGCACTCAGCCTGGGCGATAGAGACCCTGTCTCAAAAATAAATAAATAAATAAATAAATAAATAA

ATAAATAAATAAATAAAAACTAGTTCTGTGTGTATTCATTCTGTCTTTTGAGTTACATTGTTCAAGTCCTTACTTATTTTTTGACTGCTT

GACATGACACAGG

RENBPInt9.1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 193 bp

Sequence 2: ref|AC_000066.1| 193 bp

Sequence 3: ref|AC_000155.1| 193 bp

ref|NC_000023.9| CTGCACTGAGCCATGATTGTGCCACTGCACTCAGCCTGGGCGATAGAGACCCTGTCTCAA

ref|AC_000155.1| CTGCACTGAGCCATGATTGTGCCACTGCACTCAGCCTGGGCGATAGAGACCCTGTCTCAA

ref|AC_000066.1| CTGCACTGAGCCATGATTGTGCCACTGCACTCAGCCTGGGCGATAGAGACCCTGTCTCAA

************************************************************

ref|NC_000023.9| AAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAAAACTAGTTCTGTG

ref|AC_000155.1| AAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAAAACTAGTTCTGTG

ref|AC_000066.1| AAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAAAACTAGTTCTGTG

************************************************************

ref|NC_000023.9| TGTATTCATTCTGTCTTTTGAGTTACATTGTTCAAGTCCTTACTTATTTTTTGACTGCTT

ref|AC_000155.1| TGTATTCATTCTGTCTTTTGAGTTACATTGTTCAAGTCCTTACTTATTTTTTGACTGCTT

ref|AC_000066.1| TGTATTCATTCTGTCTTTTGAGTTACATTGTTCAAGTCCTTACTTATTTTTTGACTGCTT

************************************************************

ref|NC_000023.9| GACATGACACAGG

ref|AC_000155.1| GACATGACACAGG

ref|AC_000066.1| GACATGACACAGG

*************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88388(TMEM187Int1.1)	NC_000023	152892330--152892356	13.5	100	54
	AC_000066	153472848--153472874	13.5	100	54
	AC_000155	141891375--141891401	13.5	100	54

*REN88388(TMEM187Int1.1)*	Information
Amplicon	200 bp
Forward ampliprimer	GAGCAAGACCCTGTCTCAAAA
Reverse ampliprimer	ACCTCCAGGCCTTTAGCATT

>ref|NC_000023.9|

GAGCAAGACCCTGTCTCAAAAACACACACACACACACACACACACACAAAAGCAGCTGCTGGTTTCCAAGGGCCTTGCCTGGAATCAGTC

CAGAAGCTAAATTAATGTTAATTGATATTCCTTTATCCACAGAAAGACAGCAAGAAAGGTCATGCGGAGCGGGGGACATAACTAACCATA

AATGCTAAAGGCCTGGAGGT

>ref|AC_000066.1|

GAGCAAGACCCTGTCTCAAAAACACACACACACACACACACACACACAAAAGCAGCTGCTGGTTTCCAAGGGCCTTGCCTGGAATCAGTC

CAGAAGCTAAATTAATGTTAATTGATATTCCTTTATCCACAGAAAGACAGCAAGAAAGGTCATGCGGAGCGGGGGACATAACTAACCATA

AATGCTAAAGGCCTGGAGGT

>ref|AC_000155.1|

GAGCAAGACCCTGTCTCAAAAACACACACACACACACACACACACACAAAAGCAGCTGCTGGTTTCCAAGGGCCTTGCCTGGAATCAGTC

CAGAAGCTAAATTAATGTTAATTGATATTCCTTTATCCACAGAAAGACAGCAAGAAAGGTCATGCGGAGCGGGGGACATAACTAACCATA

AATGCTAAAGGCCTGGAGGT

REN88388(TMEM187Int1.1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 200 bp

Sequence 2: ref|AC_000066.1| 200 bp

Sequence 3: ref|AC_000155.1| 200 bp

ref|NC_000023.9| GAGCAAGACCCTGTCTCAAAAACACACACACACACACACACACACACAAAAGCAGCTGCT

ref|AC_000155.1| GAGCAAGACCCTGTCTCAAAAACACACACACACACACACACACACACAAAAGCAGCTGCT

ref|AC_000066.1| GAGCAAGACCCTGTCTCAAAAACACACACACACACACACACACACACAAAAGCAGCTGCT

************************************************************

ref|NC_000023.9| GGTTTCCAAGGGCCTTGCCTGGAATCAGTCCAGAAGCTAAATTAATGTTAATTGATATTC

ref|AC_000155.1| GGTTTCCAAGGGCCTTGCCTGGAATCAGTCCAGAAGCTAAATTAATGTTAATTGATATTC

ref|AC_000066.1| GGTTTCCAAGGGCCTTGCCTGGAATCAGTCCAGAAGCTAAATTAATGTTAATTGATATTC

************************************************************

ref|NC_000023.9| CTTTATCCACAGAAAGACAGCAAGAAAGGTCATGCGGAGCGGGGGACATAACTAACCATA

ref|AC_000155.1| CTTTATCCACAGAAAGACAGCAAGAAAGGTCATGCGGAGCGGGGGACATAACTAACCATA

ref|AC_000066.1| CTTTATCCACAGAAAGACAGCAAGAAAGGTCATGCGGAGCGGGGGACATAACTAACCATA

************************************************************

ref|NC_000023.9| AATGCTAAAGGCCTGGAGGT

ref|AC_000155.1| AATGCTAAAGGCCTGGAGGT

ref|AC_000066.1| AATGCTAAAGGCCTGGAGGT

********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TMEM187Int1**.2	NC_000023	152897398--152897446	9.8	100	98
**TMEM187Int1**.2	AC_000066	153477916--153477959	8.8	100	88

TMEM187Int1.2	Information
Amplicon	190-195 bp
Forward ampliprimer	GAGTGCAGTGGCGTGATCT
Reverse ampliprimer	GCGGCAGAGTGAGACTTCAT

>ref|NC_000023.9|

GAGTGCAGTGGCGTGATCTCGGCTCACTGCAACCTCCGCCTCCCGGGCTCAAGCGATTCTCCTGCCTCAG

CCTCCCGAGTAGCTGGGATTATAGGCATGTGCCACCACACCTGGCTAATTTTTTGTTTTGTTTTGTTTTG

TTTTGTTTTGTTTTGTTTTGTTTTGTTTTTCTGAGATGAAGTCTCACTCTGCCGC

>ref|AC_000066.1|

GAGTGCAGTGGCGTGATCTCGGCTCACTGCAACCTCCGCCTCCCGGGCTCAAGCGATTCTCCTGCCTCAG

CCTCCCGAGTAGCTGGGATTATAGGCATGTGCCACCACACCTGGCTAATTTTTTGTTTTGTTTTGTTTTG

TTTTGTTTTGTTTTGTTTTGTTTTTCTGAGATGAAGTCTCACTCTGCCGC

TMEM187Int1.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 195 bp

Sequence 2: ref|AC_000066.1| 190 bp

ref|NC_000023.9| GAGTGCAGTGGCGTGATCTCGGCTCACTGCAACCTCCGCCTCCCGGGCTCAAGCGATTCT

ref|AC_000066.1| GAGTGCAGTGGCGTGATCTCGGCTCACTGCAACCTCCGCCTCCCGGGCTCAAGCGATTCT

************************************************************

ref|NC_000023.9| CCTGCCTCAGCCTCCCGAGTAGCTGGGATTATAGGCATGTGCCACCACACCTGGCTAATT

ref|AC_000066.1| CCTGCCTCAGCCTCCCGAGTAGCTGGGATTATAGGCATGTGCCACCACACCTGGCTAATT

************************************************************

ref|NC_000023.9| TTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTTTCTGAGATGAA

ref|AC_000066.1| TTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTTGTTTT-----TCTGAGATGAA

******************************************** ***********

ref|NC_000023.9| GTCTCACTCTGCCGC

ref|AC_000066.1| GTCTCACTCTGCCGC

***************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88442	NC_000023	152910286--152910340	11.2	96	103
	AC_000066	153490799--153490853	11.2	96	103
	AC_000155	141909307--141909361	11.2	96	103

REN88442	Information
Amplicon	185 bp
Forward ampliprimer	CTGCCTCAGCCTCCAGAGTA
Reverse ampliprimer	GGTTGCAGTGAGCCAATAGC

>ref|NC_000023.9|

CTGCCTCAGCCTCCAGAGTAACTGGGACTACAGGTATGCACCACTGTGCCCAGCTAATTTTTTATATTTTATTTTATTTTATTTTATTTT

ATTTATTTTATTTTATTTTATTTTATTTTTGAGATGGAGTTTTGCTCTTGTCGCCCAGGCTGGAGTGCAATGCACGCTATTGGCTCACTG

CAACC

>ref|AC_000066.1|

CTGCCTCAGCCTCCAGAGTAACTGGGACTACAGGTATGCACCACTGTGCCCAGCTAATTTTTTATATTTTATTTTATTTTATTTTATTTT

ATTTATTTTATTTTATTTTATTTTATTTTTGAGATGGAGTTTTGCTCTTGTCGCCCAGGCTGGAGTGCAATGCACGCTATTGGCTCACTG

CAACC

>ref|AC_000155.1|

CTGCCTCAGCCTCCAGAGTAACTGGGACTACAGGTATGCACCACTGTGCCCAGCTAATTTTTTATATTTTATTTTATTTTATTTTATTTT

ATTTATTTTATTTTATTTTATTTTATTTTTGAGATGGAGTTTTGCTCTTGTCGCCCAGGCTGGAGTGCAATGCACGCTATTGGCTCACTG

CAACC

REN88442

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 185 bp

Sequence 2: ref|AC_000066.1| 185 bp

Sequence 3: ref|AC_000155.1| 185 bp

ref|AC_000066.1| CTGCCTCAGCCTCCAGAGTAACTGGGACTACAGGTATGCACCACTGTGCCCAGCTAATTT

ref|NC_000023.9| CTGCCTCAGCCTCCAGAGTAACTGGGACTACAGGTATGCACCACTGTGCCCAGCTAATTT

ref|AC_000155.1| CTGCCTCAGCCTCCAGAGTAACTGGGACTACAGGTATGCACCACTGTGCCCAGCTAATTT

************************************************************

ref|AC_000066.1| TTTATATTTTATTTTATTTTATTTTATTTTATTTATTTTATTTTATTTTATTTTATTTTT

ref|NC_000023.9| TTTATATTTTATTTTATTTTATTTTATTTTATTTATTTTATTTTATTTTATTTTATTTTT

ref|AC_000155.1| TTTATATTTTATTTTATTTTATTTTATTTTATTTATTTTATTTTATTTTATTTTATTTTT

************************************************************

ref|AC_000066.1| GAGATGGAGTTTTGCTCTTGTCGCCCAGGCTGGAGTGCAATGCACGCTATTGGCTCACTG

ref|NC_000023.9| GAGATGGAGTTTTGCTCTTGTCGCCCAGGCTGGAGTGCAATGCACGCTATTGGCTCACTG

ref|AC_000155.1| GAGATGGAGTTTTGCTCTTGTCGCCCAGGCTGGAGTGCAATGCACGCTATTGGCTCACTG

************************************************************

ref|AC_000066.1| CAACC

ref|NC_000023.9| CAACC

ref|AC_000155.1| CAACC

*****

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA152913.2	NC_000023	152913227--152913428	101.0	80	224
	AC_000066	153493740--153493941	101.0	80	224
	AC_000155	141912247--141912430	92.0	80	206

HEMA152913.2	Information
Amplicon	254-274 bp
Forward ampliprimer	TGAGAAGACGTGGACACTGC
Reverse ampliprimer	CCTGGGAGAAAGGGTGAGAC

>ref|NC_000023.9|

TGAGAAGACGTGGACACTGCTGTGGGCTGCAGAGGTTTGGGGGTGTGTGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGT

GTGCGTGTGTGCGTGTGTGCATGTGTGCGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGT

GCATGTGTGCATGTGTGCGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGTTTGAGACAGGGTCTCACCCTTTCTCC

CAGG

>ref|AC_000066.1|

TGAGAAGACGTGGACACTGCTGTGGGCTGCAGAGGTTTGGGGGTGTGTGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGT

GTGCGTGTGTGCGTGTGTGCATGTGTGCGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGT

GCATGTGTGCATGTGTGCGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGTTTGAGACAGGGTCTCACCCTTTCTCC

CAGG

>ref|AC_000155.1|

TGAGAAGACGTGGACACTGCTGTGGGCTGCAGAGGTTTGGGTGTGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGC

GTGTGTGCGTGTGTGCATGTGTGCGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGCATGTGTGCGT

GTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGTTTGAGACAGGGTCTCACCCTTTCTCCCAGG

HEMA152913.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 274 bp

Sequence 2: ref|AC_000066.1| 274 bp

Sequence 3: ref|AC_000155.1| 254 bp

ref|NC_000023.9| TGAGAAGACGTGGACACTGCTGTGGGCTGCAGAGGTTTGGGGGTGTGTGTGTGTGTGTGT

ref|AC_000066.1| TGAGAAGACGTGGACACTGCTGTGGGCTGCAGAGGTTTGGGGGTGTGTGTGTGTGTGTGT

ref|AC_000155.1| TGAGAAGACGTGGACACTGCTGTGGGCTGCAGAGGTTTGGG----TGTGTGTGTGTGTGT

***************************************** ***************

ref|NC_000023.9| GTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGCGTGTGTGCGTGTGTGCATGTGTGCGT

ref|AC_000066.1| GTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGCGTGTGTGCGTGTGTGCATGTGTGCGT

ref|AC_000155.1| GTGTGTGCATGTGTGTGTGTGTGTGTGTGTGTGCGTGTGTGCGTGTGTGCATGTGTGCGT

************************************************************

ref|NC_000023.9| GTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGT

ref|AC_000066.1| GTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGT

ref|AC_000155.1| GTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGTGTGTGTGTGCATGTGT--------

****************************************************

ref|NC_000023.9| GCATGTGTGCATGTGTGCGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGT

ref|AC_000066.1| GCATGTGTGCATGTGTGCGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGT

ref|AC_000155.1| --------GCATGTGTGCGTGTGTGTGTGTGTGTGTGCATGTGTGTGTGTGTGTGTGTGT

****************************************************

ref|NC_000023.9| GTGTTTGAGACAGGGTCTCACCCTTTCTCCCAGG

ref|AC_000066.1| GTGTTTGAGACAGGGTCTCACCCTTTCTCCCAGG

ref|AC_000155.1| GTGTTTGAGACAGGGTCTCACCCTTTCTCCCAGG

**********************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88627(MECP2Int2.3)	NC_000023	152955789--152955831	21.5	100	86
	AC_000066	153536300--153536322	11.5	100	46
	AC_000155	141953389--141953411	11.5	100	46

*REN88627(MECP2Int2.3)*	Information
Amplicon	205-225 bp
Forward ampliprimer	GGCGAACAAACCAGACACAT
Reverse ampliprimer	ATGCAGTAAGCCGAGATTGC

>ref|NC_000023.9|

GGCGAACAAACCAGACACATATATGTCTCAAAAAAAAATACATATATATATATATATATATATATATATATATATATATATATACACACA

CACACAAAACAAAGTTTACCTTTTTAACTATTTCTAAGTGTACAATGAATGAACTTTTTTTTTTTTTTTTTTGAGACGGAGTCTTGCTCT

GTCACCCATGCTGGGGTGTGGTGGCGCAATCTCGGCTTACTGCAT

>ref|AC_000066.1|

GGCGAACAAACCAGACACATATATGTCTCAAAAAAAAATACATATATATATATATATATATATACACACACACACAAAACAAAGTTTACC

TTTTTAACTATTTCTAAGTGTACAATGAATGAACTTTTTTTTTTTTTTTTTTGAGACGGAGTCTTGCTCTGTCACCCATGCTGGGGTGTG

GTGGCGCAATCTCGGCTTACTGCAT

>ref|AC_000155.1|

GGCGAACAAACCAGACACATATATGTCTCAAAAAAAAATACATATATATATATATATATATATACACACACACACAAAACAAAGTTTACC

TTTTTAACTATTTCTAAGTGTACAATGAATGAACTTTTTTTTTTTTTTTTTTGAGACGGAGTCTTGCTCTGTCACCCATGCTGGGGTGTG

GTGGCGCAATCTCGGCTTACTGCAT

REN88627(MECP2Int2.3)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 225 bp

Sequence 2: ref|AC_000066.1| 205 bp

Sequence 3: ref|AC_000155.1| 205 bp

ref|AC_000066.1| GGCGAACAAACCAGACACATATATGTCTCAAAAAAAAATACATATATATATATATATATA

ref|AC_000155.1| GGCGAACAAACCAGACACATATATGTCTCAAAAAAAAATACATATATATATATATATATA

ref|NC_000023.9| GGCGAACAAACCAGACACATATATGTCTCAAAAAAAAATACATATATATATATATATATA

************************************************************

ref|AC_000066.1| TATA--------------------CACACACACACAAAACAAAGTTTACCTTTTTAACTA

ref|AC_000155.1| TATA--------------------CACACACACACAAAACAAAGTTTACCTTTTTAACTA

ref|NC_000023.9| TATATATATATATATATATATATACACACACACACAAAACAAAGTTTACCTTTTTAACTA

**** ************************************

ref|AC_000066.1| TTTCTAAGTGTACAATGAATGAACTTTTTTTTTTTTTTTTTTGAGACGGAGTCTTGCTCT

ref|AC_000155.1| TTTCTAAGTGTACAATGAATGAACTTTTTTTTTTTTTTTTTTGAGACGGAGTCTTGCTCT

ref|NC_000023.9| TTTCTAAGTGTACAATGAATGAACTTTTTTTTTTTTTTTTTTGAGACGGAGTCTTGCTCT

************************************************************

ref|AC_000066.1| GTCACCCATGCTGGGGTGTGGTGGCGCAATCTCGGCTTACTGCAT

ref|AC_000155.1| GTCACCCATGCTGGGGTGTGGTGGCGCAATCTCGGCTTACTGCAT

ref|NC_000023.9| GTCACCCATGCTGGGGTGTGGTGGCGCAATCTCGGCTTACTGCAT

*********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88666/HSC270(M3)(MECP2Int2.2)	NC_000023	152964545--152964591	23.5	100	94
	AC_000066	153545036--153545080	22.5	100	90
	AC_000155	141962087--141962133	23.5	100	94

*REN88666/HSC270(M3)(MECP2Int2.2)*	Information
Amplicon	148-150 bp
Forward ampliprimer	GAATAGGCATCTGAGGACAGC
Reverse ampliprimer	TGTGCTTTGTGACATGATGG

>ref|NC_000023.9|

GAATAGGCATCTGAGGACAGCCAGAAGGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGCAGGGGGGGGGATG

GCTTCCTTCTGGTAGCCACTGCTCTAGGGAGTATTTCTCACCATCATGTCACAAAGCACA

>ref|AC_000066.1|

GAATAGGCATCTGAGGACAGCCAGAAGGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGGCAGGGGGGGGATGGC

TTCCTTCTGGTAGCCACTGCTCTAGGGAGTATTTCTCACCATCATGTCACAAAGCACA

>ref|AC_000155.1|

GAATAGGCATCTGAGGACAGCCAGAAGGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGCAGGGGGGGGATGG

CTTCCTTCTGGTAGCCACTGCTCTAGGGAGTATTTCTCACCATCATGTCACAAAGCACA

REN88666/HSC270(M3)(MECP2Int2.2)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 150 bp

Sequence 2: ref|AC_000066.1| 148 bp

Sequence 3: ref|AC_000155.1| 149 bp

ref|AC_000066.1| GAATAGGCATCTGAGGACAGCCAGAAGGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|AC_000155.1| GAATAGGCATCTGAGGACAGCCAGAAGGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|NC_000023.9| GAATAGGCATCTGAGGACAGCCAGAAGGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

************************************************************

ref|AC_000066.1| TGTGTGTGTGTGTG-GCAGGGGGGGG-ATGGCTTCCTTCTGGTAGCCACTGCTCTAGGGA

ref|AC_000155.1| TGTGTGTGTGTGTGTGCAGGGGGGGG-ATGGCTTCCTTCTGGTAGCCACTGCTCTAGGGA

ref|NC_000023.9| TGTGTGTGTGTGTGTGCAGGGGGGGGGATGGCTTCCTTCTGGTAGCCACTGCTCTAGGGA

************** *********** *********************************

ref|AC_000066.1| GTATTTCTCACCATCATGTCACAAAGCACA

ref|AC_000155.1| GTATTTCTCACCATCATGTCACAAAGCACA

ref|NC_000023.9| GTATTTCTCACCATCATGTCACAAAGCACA

******************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88881	NC_000023	153019029--153019057	14.5	100	58
	AC_000066	153599515--153599541	13.5	100	54
	AC_000155	142015871--142015897	13.5	100	54

REN88881	Information
Amplicon	141-143 bp
Forward ampliprimer	CACAGACACAGACACGCAGA
Reverse ampliprimer	TGTGTATGATTGGAATTGCCTAC

>ref|NC_000023.9|

CACAGACACAGACACGCAGACACACACAGACACACACGCACACAGAGAGAGAGAGAGAGAGAGAGAGAGAGAAACAGACCAAGAGGGCTT

TATGGATGACTTCTACCAAATCTTTAATGAGTAGGCAATTCCAATCATACACA

>ref|AC_000066.1|

CACAGACACAGACACGCAGACACACACAGACACACACGCACACAGAGAGAGAGAGAGAGAGAGAGAGAGAAACAGACCAAGAGGGCTTTA

TGGATGACTTCTACCAAATCTTTAATGAGTAGGCAATTCCAATCATACACA

>ref|AC_000155.1|

CACAGACACAGACACGCAGACACACACAGACACACACGCACACAGAGAGAGAGAGAGAGAGAGAGAGAGAAACAGACCAAGAGGGCTTTA

TGGATGACTTCTACCAAATCTTTAATGAGTAGGCAATTCCAATCATACACA

REN88881

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 143 bp

Sequence 2: ref|AC_000066.1| 141 bp

Sequence 3: ref|AC_000155.1| 141 bp

ref|AC_000066.1| CACAGACACAGACACGCAGACACACACAGACACACACGCACACAGAGAGAGAGAGAGAGA

ref|AC_000155.1| CACAGACACAGACACGCAGACACACACAGACACACACGCACACAGAGAGAGAGAGAGAGA

ref|NC_000023.9| CACAGACACAGACACGCAGACACACACAGACACACACGCACACAGAGAGAGAGAGAGAGA

************************************************************

ref|AC_000066.1| GAGAGAGAGA--AACAGACCAAGAGGGCTTTATGGATGACTTCTACCAAATCTTTAATGA

ref|AC_000155.1| GAGAGAGAGA--AACAGACCAAGAGGGCTTTATGGATGACTTCTACCAAATCTTTAATGA

ref|NC_000023.9| GAGAGAGAGAGAAACAGACCAAGAGGGCTTTATGGATGACTTCTACCAAATCTTTAATGA

********** ************************************************

ref|AC_000066.1| GTAGGCAATTCCAATCATACACA

ref|AC_000155.1| GTAGGCAATTCCAATCATACACA

ref|NC_000023.9| GTAGGCAATTCCAATCATACACA

***********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA153020.0	NC_000023	153020002--153020061	15.0	92	66
	AC_000066	153600486--153600545	15.0	92	66
	AC_000155	142016846--142016913	17.0	96	91

HEMA153020.0	Information
Amplicon	193-205 bp
Forward ampliprimer	GGAGGTGGAGGTTGCAGTAA
Reverse ampliprimer	TTTGCCCTCTCCTAACCTTTT

>ref|NC_000023.9|

GGAGGTGGAGGTTGCAGTAAGCCGAGATCGTGCCATTGCACTACAGCCTGGGTGACAGAGCGAGACTCCATCTCGAAATAGATGGATGGA

TGGATGGATGGATGGATGGATAGACAGATAGATAGATAGATAGACAGACAGACAGACAGACAGATAGATAGATAGATAAAATAAAAGGTT

AGGAGAGGGCAAA

>ref|AC_000066.1|

GGAGGTGGAGGTTGCAGTAAGCCGAGATCGTGCCATTGCACTACAGCCTGGGTGACAGAGCGAGACTCCATCTCGAAATAGATGGATGGA

TGGATGGATGGATGGATGGATAGACAGATAGATAGATAGATAGACAGACAGACAGACAGACAGATAGATAGATAGATAAAATAAAAGGTT

AGGAGAGGGCAAA

>ref|AC_000155.1|

GGAGGTGGAGGTTGCAGTAAGCCGAGATCGTGCCATTGCACTACAGCCTGGGTGACAGAGCGAGACTCCATCTCGAAATAGATGGATGGA

TGGATGGATGGATGGATGGATGGATAGATAGATAGATAGATAGATAGATAGATAGACAGACAGACAGACAGACAGATAGATAGATAGATA

AAATAAAAGGTTAGGAGAGGGCAAA

HEMA153020.0

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 193 bp

Sequence 2: ref|AC_000066.1| 193 bp

Sequence 3: ref|AC_000155.1| 205 bp

ref|NC_000023.9| GGAGGTGGAGGTTGCAGTAAGCCGAGATCGTGCCATTGCACTACAGCCTGGGTGACAGAG

ref|AC_000066.1| GGAGGTGGAGGTTGCAGTAAGCCGAGATCGTGCCATTGCACTACAGCCTGGGTGACAGAG

ref|AC_000155.1| GGAGGTGGAGGTTGCAGTAAGCCGAGATCGTGCCATTGCACTACAGCCTGGGTGACAGAG

************************************************************

ref|NC_000023.9| CGAGACTCCATCTCGAAATAGATGGATGGATGGATGGATGGATGGATGGAT---------

ref|AC_000066.1| CGAGACTCCATCTCGAAATAGATGGATGGATGGATGGATGGATGGATGGAT---------

ref|AC_000155.1| CGAGACTCCATCTCGAAATAGATGGATGGATGGATGGATGGATGGATGGATGGATAGATA

***************************************************

ref|NC_000023.9| ---AGACAGATAGATAGATAGATAGACAGACAGACAGACAGACAGATAGATAGATAGATA

ref|AC_000066.1| ---AGACAGATAGATAGATAGATAGACAGACAGACAGACAGACAGATAGATAGATAGATA

ref|AC_000155.1| GATAGATAGATAGATAGATAGATAGACAGACAGACAGACAGACAGATAGATAGATAGATA

*** *****************************************************

ref|NC_000023.9| AAATAAAAGGTTAGGAGAGGGCAAA

ref|AC_000066.1| AAATAAAAGGTTAGGAGAGGGCAAA

ref|AC_000155.1| AAATAAAAGGTTAGGAGAGGGCAAA

*************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88942	NC_000023	153034597--153034664	34.5	86	93
	AC_000066	153615068--153615131	32.5	85	85
	AC_000155	142031432--142031495	32.5	85	85

REN88942	Information
Amplicon	235 bp
Forward ampliprimer	TTGCTCCATTAGCTCCAGGT
Reverse ampliprimer	GGGCCCTTCCTTCTCTCTAA

>ref|NC_000023.9|

TTGCTCCATTAGCTCCAGGTGTTGCCATTGCAGTGTGTGTATTGTGTGTGTGTGTTTGCGTCTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGTGTGTGCACATACATTACCTGTATATTATAATGTTTAGACATCTTTTCTAAAAACTCCAAAAACGAAAACAAAAAAACTCTTTCTG

GCTGGAGAGACTGTCCTCCCAGGGCCAGCCAATTCTTAGAGAGAAGGAAGGGCCC

>ref|AC_000066.1|

TTGCTCCATTAGCTCCAGGTGTTGCCATTGCAGTGTGTGTATTGTGTGTGTGTGTTTGCGTCTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGTGCACATACATTACCTGTATATTATAATGTTTAGACATCTTTTCTAAAAACTCCAAAAACGAAAACAAAAAAACTCTTTCTGGCTG

GAGAGACTGTCCTCCCAGGGCCAGCCAATTCTTAGAGAGAAGGAAGGGCCC

>ref|AC_000155.1|

TTGCTCCATTAGCTCCAGGTGTTGCCATTGCAGTGTGTGTATTGTGTGTGTGTGTTTGCGTCTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGTGCACATACATTACCTGTATATTATAATGTTTAGACATCTTTTCTAAAAACTCCAAAAACGAAAACAAAAAAACTCTTTCTGGCTG

GAGAGACTGTCCTCCCAGGGCCAGCCAATTCTTAGAGAGAAGGAAGGGCCC

REN88942

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 235 bp

Sequence 2: ref|AC_000066.1| 231 bp

Sequence 3: ref|AC_000155.1| 231 bp

ref|AC_000066.1| TTGCTCCATTAGCTCCAGGTGTTGCCATTGCAGTGTGTGTATTGTGTGTGTGTGTTTGCG

ref|AC_000155.1| TTGCTCCATTAGCTCCAGGTGTTGCCATTGCAGTGTGTGTATTGTGTGTGTGTGTTTGCG

ref|NC_000023.9| TTGCTCCATTAGCTCCAGGTGTTGCCATTGCAGTGTGTGTATTGTGTGTGTGTGTTTGCG

************************************************************

ref|AC_000066.1| TCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG----CACATACATTACCTGTATAT

ref|AC_000155.1| TCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG----CACATACATTACCTGTATAT

ref|NC_000023.9| TCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGCACATACATTACCTGTATAT

************************************ ********************

ref|AC_000066.1| TATAATGTTTAGACATCTTTTCTAAAAACTCCAAAAACGAAAACAAAAAAACTCTTTCTG

ref|AC_000155.1| TATAATGTTTAGACATCTTTTCTAAAAACTCCAAAAACGAAAACAAAAAAACTCTTTCTG

ref|NC_000023.9| TATAATGTTTAGACATCTTTTCTAAAAACTCCAAAAACGAAAACAAAAAAACTCTTTCTG

************************************************************

ref|AC_000066.1| GCTGGAGAGACTGTCCTCCCAGGGCCAGCCAATTCTTAGAGAGAAGGAAGGGCCC

ref|AC_000155.1| GCTGGAGAGACTGTCCTCCCAGGGCCAGCCAATTCTTAGAGAGAAGGAAGGGCCC

ref|NC_000023.9| GCTGGAGAGACTGTCCTCCCAGGGCCAGCCAATTCTTAGAGAGAAGGAAGGGCCC

*******************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN88956	NC_000023	153038190--153038232	21.5	100	86
	AC_000066	153618658--153618700	21.5	100	86
	AC_000155	142035022--142035064	21.5	100	86

REN88956	Information
Amplicon	172 bp
Forward ampliprimer	CCTCAGCTCCCCAAGTAGC
Reverse ampliprimer	GCAGGCAGATTGCTTGAACT

>ref|NC_000023.9|

CCTCAGCTCCCCAAGTAGCTGGGATGACAGGCATGTGCCACCACCGGCAATTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGTCTATTATTTGTAGAGATGAGGTTTCACCATGTTAACCAGGCTGGTCTCAAACTCCTGAGTTCAAGCAATCTGCCTGC

>ref|AC_000066.1|

CCTCAGCTCCCCAAGTAGCTGGGATGACAGGCATGTGCCACCACCGGCAATTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGTCTATTATTTGTAGAGATGAGGTTTCACCATGTTAACCAGGCTGGTCTCAAACTCCTGAGTTCAAGCAATCTGCCTGC

>ref|AC_000155.1|

CCTCAGCTCCCCAAGTAGCTGGGATGACAGGCATGTGCCACCACCGGCAATTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGTCTATTATTTGTAGAGATGAGGTTTCACCATGTTAACCAGGCTGGTCTCAAACTCCTGAGTTCAAGCAATCTGCCTGC

REN88956

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 172 bp

Sequence 2: ref|AC_000066.1| 172 bp

Sequence 3: ref|AC_000155.1| 172 bp

ref|AC_000066.1| CCTCAGCTCCCCAAGTAGCTGGGATGACAGGCATGTGCCACCACCGGCAATTTGTGTGTG

ref|NC_000023.9| CCTCAGCTCCCCAAGTAGCTGGGATGACAGGCATGTGCCACCACCGGCAATTTGTGTGTG

ref|AC_000155.1| CCTCAGCTCCCCAAGTAGCTGGGATGACAGGCATGTGCCACCACCGGCAATTTGTGTGTG

************************************************************

ref|AC_000066.1| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTATTATTTGTAGAGATGAGGTTTC

ref|NC_000023.9| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTATTATTTGTAGAGATGAGGTTTC

ref|AC_000155.1| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTCTATTATTTGTAGAGATGAGGTTTC

************************************************************

ref|AC_000066.1| ACCATGTTAACCAGGCTGGTCTCAAACTCCTGAGTTCAAGCAATCTGCCTGC

ref|NC_000023.9| ACCATGTTAACCAGGCTGGTCTCAAACTCCTGAGTTCAAGCAATCTGCCTGC

ref|AC_000155.1| ACCATGTTAACCAGGCTGGTCTCAAACTCCTGAGTTCAAGCAATCTGCCTGC

****************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA153047.7	NC_000023	153047747--153047777	15.5	100	62
	NC_000023	153047777--153047824	24.0	95	87
	AC_000066	153628216--153628246	15.5	100	62
	AC_000066	153628246--153628294	25.0	91	82
	AC_000155	142044337--142044367	15.5	100	62
	AC_000155	142044367--142044422	28.0	96	103

HEMA153047.7	Information
Amplicon	272-280 bp
Forward ampliprimer	GCTCACAAGTGTAATCCCAGTG
Reverse ampliprimer	TCAAATGATCCTCCCACCTC

>ref|NC_000023.9|

GCTCACAAGTGTAATCCCAGTGATTTAGGAGGCCAAGGCAGGAAGATCCCTTGAGGCCTGGAGTTTGAAACCAGCCTGGGTAACCTAGAGAG

ACCCTGTCTCTACAATATATATATACACACACACACACACACACACACACACACATATATATATATATATATATATATATATATATATATAT

ATACATATATTTTTTTAATTAGCTGGGCATGGTGGTGTGAGCCTGTAGTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATCATTTGA

>ref|AC_000066.1|

GCTCACAAGTGTAATCCCAGTGATTTAGGAGGCCAAGGCAGGAAGATCCCTTGAGGCCTGGAGTTTGAAACCAGCCTGGGTAACCTAGAGAG

ACCCTGTCTCTACAATATATATATACACACACACACACACACACACACACACACATATATATATATATATATAATATATATATATATATATA

TATACATATATTTTTTTAATTAGCTGGGCATGGTGGTGTGAGCCTGTAGTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATCATTTGA

>ref|AC_000155.1|

GCTCACAAGTGTAATCCCAGTGATTTAGGAGGCCAAGGCAGGAAGATCCCTTGAGGCCTGGAGTTTGAAACCAGCCTGGGTAACCTAGAGAG

ACCCTGTCTCTACAATATATATATACACACACACACACACACACACACACACACATATATATATATATATATATATATATATATATATATAT

ATATATATATACATATATTTTTTTAATTAGCTGGGCATGGTGGTGTGAGCCTGTAGTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATCAT

TTGA

HEMA153047.7

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 272 bp

Sequence 2: ref|AC_000066.1| 273 bp

Sequence 3: ref|AC_000155.1| 280 bp

ref|NC_000023.9| GCTCACAAGTGTAATCCCAGTGATTTAGGAGGCCAAGGCAGGAAGATCCCTTGAGGCCTG

ref|AC_000155.1| GCTCACAAGTGTAATCCCAGTGATTTAGGAGGCCAAGGCAGGAAGATCCCTTGAGGCCTG

ref|AC_000066.1| GCTCACAAGTGTAATCCCAGTGATTTAGGAGGCCAAGGCAGGAAGATCCCTTGAGGCCTG

************************************************************

ref|NC_000023.9| GAGTTTGAAACCAGCCTGGGTAACCTAGAGAGACCCTGTCTCTACAATATATATATACAC

ref|AC_000155.1| GAGTTTGAAACCAGCCTGGGTAACCTAGAGAGACCCTGTCTCTACAATATATATATACAC

ref|AC_000066.1| GAGTTTGAAACCAGCCTGGGTAACCTAGAGAGACCCTGTCTCTACAATATATATATACAC

************************************************************

ref|NC_000023.9| ACACACACACACACACACACACACACATATATATATATATATATA-TATATATATATATA

ref|AC_000155.1| ACACACACACACACACACACACACACATATATATATATATATATA-TATATATATATATA

ref|AC_000066.1| ACACACACACACACACACACACACACATATATATATATATATATAATATATATATATATA

********************************************* **************

ref|NC_000023.9| TATATATA--------CATATATTTTTTTAATTAGCTGGGCATGGTGGTGTGAGCCTGTA

ref|AC_000155.1| TATATATATATATATACATATATTTTTTTAATTAGCTGGGCATGGTGGTGTGAGCCTGTA

ref|AC_000066.1| TATATATA--------CATATATTTTTTTAATTAGCTGGGCATGGTGGTGTGAGCCTGTA

******** ********************************************

ref|NC_000023.9| GTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATCATTTGA

ref|AC_000155.1| GTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATCATTTGA

ref|AC_000066.1| GTCCCAGCTACTCAGGAGGCTGAGGTGGGAGGATCATTTGA

*****************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA153050.0	NC_000023	153050087--153050120	17.0	100	68
	AC_000066	153630558--153630591	17.0	100	68
	AC_000155	142046686--142046719	17.0	100	68

HEMA153050.0	Information
Amplicon	286 bp
Forward ampliprimer	GTGTTGCCCAGCCTACTCTC
Reverse ampliprimer	CCTGAGAGACAGAGGTTGCAG

>ref|NC_000023.9|

GTGTTGCCCAGCCTACTCTCGAACACCTGGCCTCCAAGCAATCCTCCCATCTTGGCCTCCCAAAGTGCTGGGATTACAGGCATGAGCCCCCAC

GCCCAACCTATGCCTGATTTTTAAATTTTTACTATAAAACCTAAACTTTTAAAGCAGTAAATAATACCACTACCTGAACATCTGTGTGTGTGT

GTGTGTGTGTGTGTGTGTGTGTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGGAGTGCAGCGGCGTGATCTCGGCTCCCTGCAACCTCTGTC

TCTCAGG

>ref|AC_000066.1|

GTGTTGCCCAGCCTACTCTCGAACACCTGGCCTCCAAGCAATCCTCCCATCTTGGCCTCCCAAAGTGCTGGGATTACAGGCATGAGCCCCCAC

GCCCAACCTATGCCTGATTTTTAAATTTTTACTATAAAACCTAAACTTTTAAAGCAGTAAATAATACCACTACCTGAACATCTGTGTGTGTGT

GTGTGTGTGTGTGTGTGTGTGTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGGAGTGCAGCGGCGTGATCTCGGCTCCCTGCAACCTCTGTC

TCTCAGG

>ref|AC_000155.1|

GTGTTGCCCAGCCTACTCTCGAACACCTGGCCTCCAAGCAATCCTCCCATCTTGGCCTCCCAAAGTGCTGGGATTACAGGCATGAGCCCCCAC

GCCCAACCTATGCCTGATTTTTAAATTTTTACTATAAAACCTAAACTTTTAAAGCAGTAAATAATACCACTACCTGAACATCTGTGTGTGTGT

GTGTGTGTGTGTGTGTGTGTGTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGGAGTGCAGCGGCGTGATCTCGGCTCCCTGCAACCTCTGTC

TCTCAGG

HEMA153050.0

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 286 bp

Sequence 2: ref|AC_000066.1| 286 bp

Sequence 3: ref|AC_000155.1| 286 bp

ref|NC_000023.9| GTGTTGCCCAGCCTACTCTCGAACACCTGGCCTCCAAGCAATCCTCCCATCTTGGCCTCC

ref|AC_000155.1| GTGTTGCCCAGCCTACTCTCGAACACCTGGCCTCCAAGCAATCCTCCCATCTTGGCCTCC

ref|AC_000066.1| GTGTTGCCCAGCCTACTCTCGAACACCTGGCCTCCAAGCAATCCTCCCATCTTGGCCTCC

************************************************************

ref|NC_000023.9| CAAAGTGCTGGGATTACAGGCATGAGCCCCCACGCCCAACCTATGCCTGATTTTTAAATT

ref|AC_000155.1| CAAAGTGCTGGGATTACAGGCATGAGCCCCCACGCCCAACCTATGCCTGATTTTTAAATT

ref|AC_000066.1| CAAAGTGCTGGGATTACAGGCATGAGCCCCCACGCCCAACCTATGCCTGATTTTTAAATT

************************************************************

ref|NC_000023.9| TTTACTATAAAACCTAAACTTTTAAAGCAGTAAATAATACCACTACCTGAACATCTGTGT

ref|AC_000155.1| TTTACTATAAAACCTAAACTTTTAAAGCAGTAAATAATACCACTACCTGAACATCTGTGT

ref|AC_000066.1| TTTACTATAAAACCTAAACTTTTAAAGCAGTAAATAATACCACTACCTGAACATCTGTGT

************************************************************

ref|NC_000023.9| GTGTGTGTGTGTGTGTGTGTGTGTGTGTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGG

ref|AC_000155.1| GTGTGTGTGTGTGTGTGTGTGTGTGTGTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGG

ref|AC_000066.1| GTGTGTGTGTGTGTGTGTGTGTGTGTGTGAGATGGAGTCTCACTCTGTTGCCCAGGCTGG

************************************************************

ref|NC_000023.9| AGTGCAGCGGCGTGATCTCGGCTCCCTGCAACCTCTGTCTCTCAGG

ref|AC_000155.1| AGTGCAGCGGCGTGATCTCGGCTCCCTGCAACCTCTGTCTCTCAGG

ref|AC_000066.1| AGTGCAGCGGCGTGATCTCGGCTCCCTGCAACCTCTGTCTCTCAGG

**********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TKTL1Int1**	NC_000023	153184723--153184773	12.8	100	102
	AC_000066	153692599--153692633	8.8	100	70
	AC_000155	142108673--142108707	8.8	100	70

TKTL1Int1	Information
Amplicon	199-215 bp
Forward ampliprimer	ACACCCTGACTCGTTTCCTG
Reverse ampliprimer	CCAGGAGGTCAAGGCTGTAG

>ref|NC_000023.9|

ACACCCTGACTCGTTTCCTGCGCCAGCCTCCTACCTTGCCTTCTACTTATTAGACCCTACCTCTCATTCCCCATTTTTTAAAATTTTTAT

TTATTTATTTATTTATTTATTTATTTATTTATTTATTTATTTATTTTGAGGCAGGGTTTCACTCTGTTGCTCAGCCTAGAGGTGCAGTGG

CGTGATCATGGCTCACTACAGCCTTGACCTCCTGG

>ref|AC_000066.1|

ACACCCTGACTCGTTTCCTGCGCCAGCCTCCTACCTTGCCTTCTACTTATTAGACCCTACCTCTCATTCCCCATTTTTTAAAATTTTTAT

TTATTTATTTATTTATTTATTTATTTATTTTGAGGCAGGGTTTCACTCTGTTGCTCAGCCTAGAGGTGCAGTGGCGTGATCATGGCTCAC

TACAGCCTTGACCTCCTGG

>ref|AC_000155.1|

ACACCCTGACTCGTTTCCTGCGCCAGCCTCCTACCTTGCCTTCTACTTATTAGACCCTACCTCTCATTCCCCATTTTTTAAAATTTTTAT

TTATTTATTTATTTATTTATTTATTTATTTTGAGGCAGGGTTTCACTCTGTTGCTCAGCCTAGAGGTGCAGTGGCGTGATCATGGCTCAC

TACAGCCTTGACCTCCTGG

TKTL1Int1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 215 bp

Sequence 2: ref|AC_000066.1| 199 bp

Sequence 3: ref|AC_000155.1| 199 bp

ref|AC_000066.1| ACACCCTGACTCGTTTCCTGCGCCAGCCTCCTACCTTGCCTTCTACTTATTAGACCCTAC

ref|AC_000155.1| ACACCCTGACTCGTTTCCTGCGCCAGCCTCCTACCTTGCCTTCTACTTATTAGACCCTAC

ref|NC_000023.9| ACACCCTGACTCGTTTCCTGCGCCAGCCTCCTACCTTGCCTTCTACTTATTAGACCCTAC

************************************************************

ref|AC_000066.1| CTCTCATTCCCCATTTTTTAAAATTTTTATTTATTTATTTATTTATTTATTTATTTATTT

ref|AC_000155.1| CTCTCATTCCCCATTTTTTAAAATTTTTATTTATTTATTTATTTATTTATTTATTTATTT

ref|NC_000023.9| CTCTCATTCCCCATTTTTTAAAATTTTTATTTATTTATTTATTTATTTATTTATTTATTT

************************************************************

ref|AC_000066.1| ----------------TGAGGCAGGGTTTCACTCTGTTGCTCAGCCTAGAGGTGCAGTGG

ref|AC_000155.1| ----------------TGAGGCAGGGTTTCACTCTGTTGCTCAGCCTAGAGGTGCAGTGG

ref|NC_000023.9| ATTTATTTATTTATTTTGAGGCAGGGTTTCACTCTGTTGCTCAGCCTAGAGGTGCAGTGG

********************************************

ref|AC_000066.1| CGTGATCATGGCTCACTACAGCCTTGACCTCCTGG

ref|AC_000155.1| CGTGATCATGGCTCACTACAGCCTTGACCTCCTGG

ref|NC_000023.9| CGTGATCATGGCTCACTACAGCCTTGACCTCCTGG

***********************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN89483(TAZInt5)	NC_000023	153296967--153297133	41.0	82	95
	AC_000066	153804830--153804996	41.0	84	102
	AC_000155	142221162--142221328	41.0	84	102

*REN89483(TAZInt5)*	Information
Amplicon	319-320 bp
Forward ampliprimer	AGGGCTGGTCTTGAACTCCT
Reverse ampliprimer	CACCCCACCTGGCTATCTAA

>ref|NC_000023.9|

AGGGCTGGTCTTGAACTCCTAGACTCAAGCGATCCTCCCGCCTCAGCCTCTCAAGTGCTGGGATCACAGGCATGAGCCATTGTGCCCAAC

CCAGCCCAGGAATTTGAGGCCAGCCTGAGCAATATAGTGAGACCCCAGGTATAGATAGATAGGTAGATAGATAGGTAGGTAGGTAGGTAG

ATAGATAGATACGTAGGTGGGTAGGTAGGTGGGTAGATAGGTAGGTAGGTACGTAGGTAGATAGATAGATAGATACATACATACATAGAT

AGATAAGATAGATAGATTGATTAGATAGATTAGATAGCCAGGTGGGGTG

>ref|AC_000066.1|

AGGGCTGGTCTTGAACTCCTAGACTCAAGCGATCCTCCCGCCTCAGCCTCTCAAAGTGCTGGGATCACAGGCATGAGCCATTGTGCCCAA

CCCAGCCCAGGAATTTGAGGCCAGCCTGAGCAATATAGTGAGACCCCAGGTATAGATAGATAGGTAGATAGATAGGTAGGTAGGTAGGTA

GATAGATAGATAGGTAGGTGGGTAGGTAGGTGGGTAGATAGGTAGGTAGGTACGTAGGTAGATAGATAGATAGATACATACATACATAGA

TAGATAAGATAGATAGATTGATTAGATAGATTAGATAGCCAGGTGGGGTG

>ref|AC_000155.1|

AGGGCTGGTCTTGAACTCCTAGACTCAAGCGATCCTCCCGCCTCAGCCTCTCAAAGTGCTGGGATCACAGGCATGAGCCATTGTGCCCAA

CCCAGCCCAGGAATTTGAGGCCAGCCTGAGCAATATAGTGAGACCCCAGGTATAGATAGATAGGTAGATAGATAGGTAGGTAGGTAGGTA

GATAGATAGATAGGTAGGTGGGTAGGTAGGTGGGTAGATAGGTAGGTAGGTACGTAGGTAGATAGATAGATAGATACATACATACATAGA

TAGATAAGATAGATAGATTGATTAGATAGATTAGATAGCCAGGTGGGGTG

REN89483(TAZInt5)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 319 bp

Sequence 2: ref|AC_000066.1| 320 bp

Sequence 3: ref|AC_000155.1| 320 bp

ref|AC_000066.1| AGGGCTGGTCTTGAACTCCTAGACTCAAGCGATCCTCCCGCCTCAGCCTCTCAAAGTGCT

ref|AC_000155.1| AGGGCTGGTCTTGAACTCCTAGACTCAAGCGATCCTCCCGCCTCAGCCTCTCAAAGTGCT

ref|NC_000023.9| AGGGCTGGTCTTGAACTCCTAGACTCAAGCGATCCTCCCGCCTCAGCCTCTCAA-GTGCT

****************************************************** *****

ref|AC_000066.1| GGGATCACAGGCATGAGCCATTGTGCCCAACCCAGCCCAGGAATTTGAGGCCAGCCTGAG

ref|AC_000155.1| GGGATCACAGGCATGAGCCATTGTGCCCAACCCAGCCCAGGAATTTGAGGCCAGCCTGAG

ref|NC_000023.9| GGGATCACAGGCATGAGCCATTGTGCCCAACCCAGCCCAGGAATTTGAGGCCAGCCTGAG

************************************************************

ref|AC_000066.1| CAATATAGTGAGACCCCAGGTATAGATAGATAGGTAGATAGATAGGTAGGTAGGTAGGTA

ref|AC_000155.1| CAATATAGTGAGACCCCAGGTATAGATAGATAGGTAGATAGATAGGTAGGTAGGTAGGTA

ref|NC_000023.9| CAATATAGTGAGACCCCAGGTATAGATAGATAGGTAGATAGATAGGTAGGTAGGTAGGTA

************************************************************

ref|AC_000066.1| GATAGATAGATAGGTAGGTGGGTAGGTAGGTGGGTAGATAGGTAGGTAGGTACGTAGGTA

ref|AC_000155.1| GATAGATAGATAGGTAGGTGGGTAGGTAGGTGGGTAGATAGGTAGGTAGGTACGTAGGTA

ref|NC_000023.9| GATAGATAGATACGTAGGTGGGTAGGTAGGTGGGTAGATAGGTAGGTAGGTACGTAGGTA

************ ***********************************************

ref|AC_000066.1| GATAGATAGATAGATACATACATACATAGATAGATAAGATAGATAGATTGATTAGATAGA

ref|AC_000155.1| GATAGATAGATAGATACATACATACATAGATAGATAAGATAGATAGATTGATTAGATAGA

ref|NC_000023.9| GATAGATAGATAGATACATACATACATAGATAGATAAGATAGATAGATTGATTAGATAGA

************************************************************

ref|AC_000066.1| TTAGATAGCCAGGTGGGGTG

ref|AC_000155.1| TTAGATAGCCAGGTGGGGTG

ref|NC_000023.9| TTAGATAGCCAGGTGGGGTG

********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA153333.1	NC_000023	153333110--153333151	10.8	89	68
	AC_000066	153840959--153841000	10.8	89	68
	AC_000155	142257523--142257564	10.8	89	68

HEMA153333.1	Information
Amplicon	182 bp
Forward ampliprimer	CACCTGGCGTCACTAGGAAT
Reverse ampliprimer	CTGGGAGACAGAGCAAGACC

>ref|NC_000023.9|

CACCTGGCGTCACTAGGAATAGAGATGAGAGTAGACTAATGTTTTCTTTCTTTTCTTTTCCTTCCCTCCTTCCTTCCTTCTTCCTTCCTT

CCTTCCTTCTTTCTTCCCTCCCCTCCCCTCCCCTCTCCTTCCTTTCCTCTTTCTTTTTTTTTTTTGAGACTGGGTCTTGCTCTGTCTCCC

>ref|AC_000066.1|

CACCTGGCGTCACTAGGAATAGAGATGAGAGTAGACTAATGTTTTCTTTCTTTTCTTTTCCTTCCCTCCTTCCTTCCTTCTTCCTTCCTT

CCTTCCTTCTTTCTTCCCTCCCCTCCCCTCCCCTCTCCTTCCTTTCCTCTTTCTTTTTTTTTTTTGAGACTGGGTCTTGCTCTGTCTCCC

>ref|AC_000155.1|

CACCTGGCGTCACTAGGAATAGAGATGAGAGTAGACTAATGTTTTCTTTCTTTTCTTTTCCTTCCCTCCTTCCTTCCTTCTTCCTTCCTT

CCTTCCTTCTTTCTTCCCTCCCCTCCCCTCCCCTCTCCTTCCTTTCCTCTTTCTTTTTTTTTTTTGAGACTGGGTCTTGCTCTGTCTCCC

HEMA153333.1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 182 bp

Sequence 2: ref|AC_000066.1| 182 bp

Sequence 3: ref|AC_000155.1| 182 bp

ref|NC_000023.9| CACCTGGCGTCACTAGGAATAGAGATGAGAGTAGACTAATGTTTTCTTTCTTTTCTTTTC

ref|AC_000155.1| CACCTGGCGTCACTAGGAATAGAGATGAGAGTAGACTAATGTTTTCTTTCTTTTCTTTTC

ref|AC_000066.1| CACCTGGCGTCACTAGGAATAGAGATGAGAGTAGACTAATGTTTTCTTTCTTTTCTTTTC

************************************************************

ref|NC_000023.9| CTTCCCTCCTTCCTTCCTTCTTCCTTCCTTCCTTCCTTCTTTCTTCCCTCCCCTCCCCTC

ref|AC_000155.1| CTTCCCTCCTTCCTTCCTTCTTCCTTCCTTCCTTCCTTCTTTCTTCCCTCCCCTCCCCTC

ref|AC_000066.1| CTTCCCTCCTTCCTTCCTTCTTCCTTCCTTCCTTCCTTCTTTCTTCCCTCCCCTCCCCTC

************************************************************

ref|NC_000023.9| CCCTCTCCTTCCTTTCCTCTTTCTTTTTTTTTTTTGAGACTGGGTCTTGCTCTGTCTCCC

ref|AC_000155.1| CCCTCTCCTTCCTTTCCTCTTTCTTTTTTTTTTTTGAGACTGGGTCTTGCTCTGTCTCCC

ref|AC_000066.1| CCCTCTCCTTCCTTTCCTCTTTCTTTTTTTTTTTTGAGACTGGGTCTTGCTCTGTCTCCC

************************************************************

ref|NC_000023.9| AG

ref|AC_000155.1| AG

ref|AC_000066.1| AG

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
G65800	NC_000023	153376711--153376758	12.3	91	80
	AC_000066	153884568--153884615	12.3	91	80
	AC_000155	142300522--142300569	12.3	91	80

G65800	Information
Amplicon	168 bp
Forward ampliprimer	TCCAAAGTGCTGGGATTACA
Reverse ampliprimer	AGGGTGCAGTGAGCTGAGAT

>ref|NC_000023.9|

TCCAAAGTGCTGGGATTACAAACATGAGCCATTGCAGCAGGCTGACAAAGGTGTTTTTTGTTTGTTTGTTTGTTTGTTTGTTTGTCTGTT

TGTTTGTTTTTGAGATGGAGTTTGGCTCTTGTTGCCCAGGCTGGAGTGCAATGGCGCGATCTCAGCTCACTGCACCCT

>ref|AC_000066.1|

TCCAAAGTGCTGGGATTACAAACATGAGCCATTGCAGCAGGCTGACAAAGGTGTTTTTTGTTTGTTTGTTTGTTTGTTTGTTTGTCTGTT

TGTTTGTTTTTGAGATGGAGTTTGGCTCTTGTTGCCCAGGCTGGAGTGCAATGGCGCGATCTCAGCTCACTGCACCCT

>ref|AC_000155.1|

TCCAAAGTGCTGGGATTACAAACATGAGCCATTGCAGCAGGCTGACAAAGGTGTTTTTTGTTTGTTTGTTTGTTTGTTTGTTTGTCTGTT

TGTTTGTTTTTGAGATGGAGTTTGGCTCTTGTTGCCCAGGCTGGAGTGCAATGGCGCGATCTCAGCTCACTGCACCCT

G65800

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 168 bp

Sequence 2: ref|AC_000066.1| 168 bp

Sequence 3: ref|AC_000155.1| 168 bp

ref|AC_000066.1| TCCAAAGTGCTGGGATTACAAACATGAGCCATTGCAGCAGGCTGACAAAGGTGTTTTTTG

ref|NC_000023.9| TCCAAAGTGCTGGGATTACAAACATGAGCCATTGCAGCAGGCTGACAAAGGTGTTTTTTG

ref|AC_000155.1| TCCAAAGTGCTGGGATTACAAACATGAGCCATTGCAGCAGGCTGACAAAGGTGTTTTTTG

************************************************************

ref|AC_000066.1| TTTGTTTGTTTGTTTGTTTGTTTGTCTGTTTGTTTGTTTTTGAGATGGAGTTTGGCTCTT

ref|NC_000023.9| TTTGTTTGTTTGTTTGTTTGTTTGTCTGTTTGTTTGTTTTTGAGATGGAGTTTGGCTCTT

ref|AC_000155.1| TTTGTTTGTTTGTTTGTTTGTTTGTCTGTTTGTTTGTTTTTGAGATGGAGTTTGGCTCTT

************************************************************

ref|AC_000066.1| GTTGCCCAGGCTGGAGTGCAATGGCGCGATCTCAGCTCACTGCACCCT

ref|NC_000023.9| GTTGCCCAGGCTGGAGTGCAATGGCGCGATCTCAGCTCACTGCACCCT

ref|AC_000155.1| GTTGCCCAGGCTGGAGTGCAATGGCGCGATCTCAGCTCACTGCACCCT

************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN89870(FAM3AInt2)	NC_000023	153393770--153393811	14.0	100	84
	AC_000066	153901632--153901664	11.0	100	66
	AC_000155	142317343--142317375	11.0	100	66

*REN89870(FAM3AInt2)*	Information
Amplicon	233-236 bp
Forward ampliprimer	TCATGCCTGTAGCACTTTGG
Reverse ampliprimer	CCTGGGTTCAAGCGATTCTA

>ref|NC_000023.9|

TCATGCCTGTAGCACTTTGGGAGGCTGAGGCGGGTAGATTGCTTGAGCCCAGGAGTTTGAGACCAGCCTGGGCAACATGGTGAAACCCCA

TCTCTACTAAAAATACAATAATAATAATAATAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGCCGGGTTTGGTGGTGCG

CACCTGTAGTCTCAGCTACTCGGGAGGCTGGGGCACTAGAATCGCTTGAACCCAGG

>ref|AC_000066.1|

TCATGCCTGTAGCACTTTGGGAGGCTGAGGCGGGTAGATTGCTTGAGCCCAGGAGTTTGAGACCAGCCTGGGCAACATGGTGAAACCCCA

TCTCTACTAAAAATACAATAATAATAATAATAATAATAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGCCGGGTTTGGTGGTGCGCAC

CTGTAGTCTCAGCTACTCGGGAGGCTGGGGCACTAGAATCGCTTGAACCCAGG

>ref|AC_000155.1|

TCATGCCTGTAGCACTTTGGGAGGCTGAGGCGGGTAGATTGCTTGAGCCCAGGAGTTTGAGACCAGCCTGGGCAACATGGTGAAACCCCA

TCTCTACTAAAAATACAATAATAATAATAATAATAATAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGCCGGGTTTGGTGGTGCGCAC

CTGTAGTCTCAGCTACTCGGGAGGCTGGGGCACTAGAATCGCTTGAACCCAGG

REN89870(FAM3AInt2)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 236 bp

Sequence 2: ref|AC_000066.1| 233 bp

Sequence 3: ref|AC_000155.1| 233 bp

ref|AC_000066.1|      TCATGCCTGTAGCACTTTGGGAGGCTGAGGCGGGTAGATTGCTTGAGCCCAGGAGTTTGA

ref|AC_000155.1|      TCATGCCTGTAGCACTTTGGGAGGCTGAGGCGGGTAGATTGCTTGAGCCCAGGAGTTTGA

ref|NC_000023.9|      TCATGCCTGTAGCACTTTGGGAGGCTGAGGCGGGTAGATTGCTTGAGCCCAGGAGTTTGA

                      ************************************************************

ref|AC_000066.1|      GACCAGCCTGGGCAACATGGTGAAACCCCATCTCTACTAAAAATACAATAATAATAATAA

ref|AC_000155.1|      GACCAGCCTGGGCAACATGGTGAAACCCCATCTCTACTAAAAATACAATAATAATAATAA

ref|NC_000023.9|      GACCAGCCTGGGCAACATGGTGAAACCCCATCTCTACTAAAAATACAATAATAATAATAA

                      ************************************************************

ref|AC_000066.1|      TAATAATAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAG---CCGGGTTTGGTGGTGCG

ref|AC_000155.1|      TAATAATAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAG---CCGGGTTTGGTGGTGCG

ref|NC_000023.9|      TAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGAAGCCGGGTTTGGTGGTGCG

                      *** ** *********************************   *****************

ref|AC_000066.1|      CACCTGTAGTCTCAGCTACTCGGGAGGCTGGGGCACTAGAATCGCTTGAACCCAGG

ref|AC_000155.1|      CACCTGTAGTCTCAGCTACTCGGGAGGCTGGGGCACTAGAATCGCTTGAACCCAGG

ref|NC_000023.9|      CACCTGTAGTCTCAGCTACTCGGGAGGCTGGGGCACTAGAATCGCTTGAACCCAGG

                      ********************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN89898/REN89899	NC_000023	153401350--153401379	15.0	100	60
	AC_000066	153909203--153909232	15.0	100	60
	AC_000155	142324906--142324937	16.0	100	64

New!

WARNING!

This microsatellite marker was firstly predicted In-Silico as informative ^[1], and it has recently been tried-and-tested, apparently (no primer sequences provided), using a pair set of ampliprimers different to the one here provided ^[2]. The authors reported that genotyping of this STR marker yielded peak patterns that made allele calling difficult and, therefore, excluded it form the multiplex assay. Worth of note, the latter study also reported similar impairing effects for two other STR marker loci (REN90371 and stSG604469), all of which contain a [AT] repeat unit.

^[1]

^[2]

REN89898/REN89899	Information
Amplicon	120-122 bp
Forward ampliprimer	GATTGCTTGAAGCCAGGAAC
Reverse ampliprimer	TGTGGAGTCTAGCTGCCCTTT

>ref|NC_000023.9|

GATTGCTTGAAGCCAGGAACTTGAGACCAGCCTGGGCAACAAAGTGAGACCCTGCATATATATATATATATATATATATATATATTTACA

CAAATAAATAAAGGGCAGCTAGACTCCACA

>ref|AC_000066.1|

GATTGCTTGAAGCCAGGAACTTGAGACCAGCCTGGGCAACAAAGTGAGACCCTGCATATATATATATATATATATATATATATATTTACA

CAAATAAATAAAGGGCAGCTAGACTCCACA

>ref|AC_000155.1|

GATTGCTTGAAGCCAGGAACTTGAGACCAGCCTGGGCAACAAAGTGAGACCCTGCATATATATATATATATATATATATATATATATTTA

CACAAATAAATAAAGGGCAGCTAGACTCCACA

REN89898/REN89899

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 120 bp

Sequence 2: ref|AC_000066.1| 120 bp

Sequence 3: ref|AC_000155.1| 122 bp

ref|NC_000023.9| GATTGCTTGAAGCCAGGAACTTGAGACCAGCCTGGGCAACAAAGTGAGACCCTGCATATA

ref|AC_000066.1| GATTGCTTGAAGCCAGGAACTTGAGACCAGCCTGGGCAACAAAGTGAGACCCTGCATATA

ref|AC_000155.1| GATTGCTTGAAGCCAGGAACTTGAGACCAGCCTGGGCAACAAAGTGAGACCCTGCATATA

************************************************************

ref|NC_000023.9| TATATATATATATATATATATATAT--TTACACAAATAAATAAAGGGCAGCTAGACTCCA

ref|AC_000066.1| TATATATATATATATATATATATAT--TTACACAAATAAATAAAGGGCAGCTAGACTCCA

ref|AC_000155.1| TATATATATATATATATATATATATATTTACACAAATAAATAAAGGGCAGCTAGACTCCA

************************* *********************************

ref|NC_000023.9| CA

ref|AC_000066.1| CA

ref|AC_000155.1| CA

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604115/REN89922	NC_000023	153408151--153408189	19.5	89	60
	AC_000066	153916004--153916042	19.5	89	60
	AC_000155	142331539--142331577	19.5	89	60

stSG604115/REN89922	Information
Amplicon	211 bp
Forward ampliprimer	GGCAACTTGGGGGTTAATTT
Reverse ampliprimer	AGGAGACTGAGGTGGGAGGT

>ref|NC_000023.9|

GGCAACTTGGGGGTTAATTTGTATCATGGGGTCCTAGTTTTCTCTGAAAATTTGGCCTTGCAAATGTGTGTGTGTGTGTGTGTGTATGAG

TGTGTGTGTGTGTATATATATATATTTATATATATATTTGAGACAGGGTCTTGCTGTGTGGCCCAGGCTGGAGTGCAATGGTGTGATCAT

GGCCCAAGTGAACCTCCCACCTCAGTCTCCT

>ref|AC_000066.1|

GGCAACTTGGGGGTTAATTTGTATCATGGGGTCCTAGTTTTCTCTGAAAATTTGGCCTTGCAAATGTGTGTGTGTGTGTGTGTGTATGAG

TGTGTGTGTGTGTATATATATATATTTATATATATATTTGAGACAGGGTCTTGCTGTGTGGCCCAGGCTGGAGTGCAATGGTGTGATCAT

GGCCCAAGTGAACCTCCCACCTCAGTCTCCT

>ref|AC_000155.1

GGCAACTTGGGGGTTAATTTGTATCATGGGGTCCTAGTTTTCTCTGAAAATTTGGCCTTGCAAATGTGTGTGTGTGTGTGTGTGTATGAG

TGTGTGTGTGTGTATATATATATATTTATATATATATTTGAGACAGGGTCTTGCTGTGTGGCCCAGGCTGGAGTGCAATGGTGTGATCAT

GGCCCAAGTGAACCTCCCACCTCAGTCTCCT

stSG604115/REN89922

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 211 bp

Sequence 2: ref|AC_000066.1| 211 bp

Sequence 3: ref|AC_000155.1| 211 bp

ref|AC_000066.1| GGCAACTTGGGGGTTAATTTGTATCATGGGGTCCTAGTTTTCTCTGAAAATTTGGCCTTG

ref|NC_000023.9| GGCAACTTGGGGGTTAATTTGTATCATGGGGTCCTAGTTTTCTCTGAAAATTTGGCCTTG

ref|AC_000155.1| GGCAACTTGGGGGTTAATTTGTATCATGGGGTCCTAGTTTTCTCTGAAAATTTGGCCTTG

************************************************************

ref|AC_000066.1| CAAATGTGTGTGTGTGTGTGTGTGTATGAGTGTGTGTGTGTGTATATATATATATTTATA

ref|NC_000023.9| CAAATGTGTGTGTGTGTGTGTGTGTATGAGTGTGTGTGTGTGTATATATATATATTTATA

ref|AC_000155.1| CAAATGTGTGTGTGTGTGTGTGTGTATGAGTGTGTGTGTGTGTATATATATATATTTATA

************************************************************

ref|AC_000066.1| TATATATTTGAGACAGGGTCTTGCTGTGTGGCCCAGGCTGGAGTGCAATGGTGTGATCAT

ref|NC_000023.9| TATATATTTGAGACAGGGTCTTGCTGTGTGGCCCAGGCTGGAGTGCAATGGTGTGATCAT

ref|AC_000155.1| TATATATTTGAGACAGGGTCTTGCTGTGTGGCCCAGGCTGGAGTGCAATGGTGTGATCAT

************************************************************

ref|AC_000066.1| GGCCCAAGTGAACCTCCCACCTCAGTCTCCT

ref|NC_000023.9| GGCCCAAGTGAACCTCCCACCTCAGTCTCCT

ref|AC_000155.1| GGCCCAAGTGAACCTCCCACCTCAGTCTCCT

*******************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90010/GDB:179459(IKBKGInt1)	NC_000023	153431644--153431687	22.0	95	79
	AC_000066	153939499--153939542	22.0	95	79
	AC_000155	142354960--142355003	22.0	95	79

New!	This repeat element was previously reported by others as located on G6PD; we reassigned it to the IKBKG locus^[1].
^[1]	Machado FB, Medina-Acosta E. An IKBKG, and not a G6PD, short tandem repeat marker is used in indirect diagnosis of haemophilia A. Haemophilia 2008, 14:849-850.

*REN90010/GDB:179459(IKBKGInt1)*	Information
Amplicon	183 bp
Forward ampliprimer	ATCGGTTGGGGGATACATTT
Reverse ampliprimer	TCTACCCAAGGCCTTCTTTG

>ref|NC_000023.9|

ATCGGTTGGGGGATACATTTTAACAGGATTTCTTGATGGATTAGATGTGCTCTTGTGTGTGTGTGTGTGTGTGTGTGTATGTGTGTGTGT

GTGTGTGAGAGAGAGAGAGAGAATCAAAAAAGACTTTTGACAGCTGGAGGATGGATTCGCCATCAGCTGATAACAAAGAAGGCCTTGGGT

AGA

>ref|AC_000066.1|

ATCGGTTGGGGGATACATTTTAACAGGATTTCTTGATGGATTAGATGTGCTCTTGTGTGTGTGTGTGTGTGTGTGTGTATGTGTGTGTGT

GTGTGTGAGAGAGAGAGAGAGAATCAAAAAAGACTTTTGACAGCTGGAGGATGGATTCGCCATCAGCTGATAACAAAGAAGGCCTTGGGT

AGA

>ref|AC_000155.1|

ATCGGTTGGGGGATACATTTTAACAGGATTTCTTGATGGATTAGATGTGCTCTTGTGTGTGTGTGTGTGTGTGTGTGTATGTGTGTGTGT

GTGTGTGAGAGAGAGAGAGAGAATCAAAAAAGACTTTTGACAGCTGGAGGATGGATTCGCCATCAGCTGATAACAAAGAAGGCCTTGGGT

AGA

REN90010/GDB:179459(IKBKGInt1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 183 bp

Sequence 2: ref|AC_000066.1| 183 bp

Sequence 3: ref|AC_000155.1| 183 bp

ref|AC_000066.1| ATCGGTTGGGGGATACATTTTAACAGGATTTCTTGATGGATTAGATGTGCTCTTGTGTGT

ref|NC_000023.9| ATCGGTTGGGGGATACATTTTAACAGGATTTCTTGATGGATTAGATGTGCTCTTGTGTGT

ref|AC_000155.1| ATCGGTTGGGGGATACATTTTAACAGGATTTCTTGATGGATTAGATGTGCTCTTGTGTGT

************************************************************

ref|AC_000066.1| GTGTGTGTGTGTGTGTGTATGTGTGTGTGTGTGTGTGAGAGAGAGAGAGAGAATCAAAAA

ref|NC_000023.9| GTGTGTGTGTGTGTGTGTATGTGTGTGTGTGTGTGTGAGAGAGAGAGAGAGAATCAAAAA

ref|AC_000155.1| GTGTGTGTGTGTGTGTGTATGTGTGTGTGTGTGTGTGAGAGAGAGAGAGAGAATCAAAAA

************************************************************

ref|AC_000066.1| AGACTTTTGACAGCTGGAGGATGGATTCGCCATCAGCTGATAACAAAGAAGGCCTTGGGT

ref|NC_000023.9| AGACTTTTGACAGCTGGAGGATGGATTCGCCATCAGCTGATAACAAAGAAGGCCTTGGGT

ref|AC_000155.1| AGACTTTTGACAGCTGGAGGATGGATTCGCCATCAGCTGATAACAAAGAAGGCCTTGGGT

************************************************************

ref|AC_000066.1| AGA

ref|NC_000023.9| AGA

ref|AC_000155.1| AGA

***

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604170/REN90051/REN90052	NC_000023	153477086--153477279	103.0	81	225
	AC_000066	153983899--153984092	103.0	81	225
	AC_000155	142369096--142369289	103.0	81	225

stSG604170/REN90051/REN90052	Information
Amplicon	318 bp
Forward ampliprimer	ATGGGCTTGGACTTGGTATG
Reverse ampliprimer	AGCAGCTCCTCACCAGGAT

>ref|NC_000023.9|

ATGGGCTTGGACTTGGTATGTCTTTTTCAGTTAATTGTATGATAAATTTTATTGTTTTAACAAAAGTACCAGTCCATGATAGATTGGAAC

TATGAGAGAGATACAGAGACAGAGAGAGAGAGGAGAGAGAAGAGAGAGAGGAGAGAGAAAAGAGAGAGCAGAGAGAGAGAAGAGAGAGAA

GAGAGGAGAGAGAAAAGAGAGAGGAGAGAGAGAGAGGAGAGAGAGAGAGAGAGAGGAGAGGGAGAGAGAGAGAGAGGAGAGACAGAGAGA

GAGAGGAGAGAGAGAGAAATTCCTCAGAGATCCTGGTGAGGAGCTGCT

>ref|AC_000066.1|

ATGGGCTTGGACTTGGTATGTCTTTTTCAGTTAATTGTATGATAAATTTTATTGTTTTAACAAAAGTACCAGTCCATGATAGATTGGAAC

TATGAGAGAGATACAGAGACAGAGAGAGAGAGGAGAGAGAAGAGAGAGAGGAGAGAGAAAAGAGAGAGCAGAGAGAGAGAAGAGAGAGAA

GAGAGGAGAGAGAAAAGAGAGAGGAGAGAGAGAGAGGAGAGAGAGAGAGAGAGAGGAGAGGGAGAGAGAGAGAGAGGAGAGACAGAGAGA

GAGAGGAGAGAGAGAGAAATTCCTCAGAGATCCTGGTGAGGAGCTGCT

>ref|AC_000155.1|

ATGGGCTTGGACTTGGTATGTCTTTTTCAGTTAATTGTATGATAAATTTTATTGTTTTAACAAAAGTACCAGTCCATGATAGATTGGAAC

TATGAGAGAGATACAGAGACAGAGAGAGAGAGGAGAGAGAAGAGAGAGAGGAGAGAGAAAAGAGAGAGCAGAGAGAGAGAAGAGAGAGAA

GAGAGGAGAGAGAAAAGAGAGAGGAGAGAGAGAGAGGAGAGAGAGAGAGAGAGAGGAGAGGGAGAGAGAGAGAGAGGAGAGACAGAGAGA

GAGAGGAGAGAGAGAGAAATTCCTCAGAGATCCTGGTGAGGAGCTGCT

stSG604170/REN90051/REN90052

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 318 bp

Sequence 2: ref|AC_000066.1| 318 bp

Sequence 3: ref|AC_000155.1| 318 bp

ref|AC_000066.1| ATGGGCTTGGACTTGGTATGTCTTTTTCAGTTAATTGTATGATAAATTTTATTGTTTTAA

ref|NC_000023.9| ATGGGCTTGGACTTGGTATGTCTTTTTCAGTTAATTGTATGATAAATTTTATTGTTTTAA

ref|AC_000155.1| ATGGGCTTGGACTTGGTATGTCTTTTTCAGTTAATTGTATGATAAATTTTATTGTTTTAA

************************************************************

ref|AC_000066.1| CAAAAGTACCAGTCCATGATAGATTGGAACTATGAGAGAGATACAGAGACAGAGAGAGAG

ref|NC_000023.9| CAAAAGTACCAGTCCATGATAGATTGGAACTATGAGAGAGATACAGAGACAGAGAGAGAG

ref|AC_000155.1| CAAAAGTACCAGTCCATGATAGATTGGAACTATGAGAGAGATACAGAGACAGAGAGAGAG

************************************************************

ref|AC_000066.1| AGGAGAGAGAAGAGAGAGAGGAGAGAGAAAAGAGAGAGCAGAGAGAGAGAAGAGAGAGAA

ref|NC_000023.9| AGGAGAGAGAAGAGAGAGAGGAGAGAGAAAAGAGAGAGCAGAGAGAGAGAAGAGAGAGAA

ref|AC_000155.1| AGGAGAGAGAAGAGAGAGAGGAGAGAGAAAAGAGAGAGCAGAGAGAGAGAAGAGAGAGAA

************************************************************

ref|AC_000066.1| GAGAGGAGAGAGAAAAGAGAGAGGAGAGAGAGAGAGGAGAGAGAGAGAGAGAGAGGAGAG

ref|NC_000023.9| GAGAGGAGAGAGAAAAGAGAGAGGAGAGAGAGAGAGGAGAGAGAGAGAGAGAGAGGAGAG

ref|AC_000155.1| GAGAGGAGAGAGAAAAGAGAGAGGAGAGAGAGAGAGGAGAGAGAGAGAGAGAGAGGAGAG

************************************************************

ref|AC_000066.1| GGAGAGAGAGAGAGAGGAGAGACAGAGAGAGAGAGGAGAGAGAGAGAAATTCCTCAGAGA

ref|NC_000023.9| GGAGAGAGAGAGAGAGGAGAGACAGAGAGAGAGAGGAGAGAGAGAGAAATTCCTCAGAGA

ref|AC_000155.1| GGAGAGAGAGAGAGAGGAGAGACAGAGAGAGAGAGGAGAGAGAGAGAAATTCCTCAGAGA

************************************************************

ref|AC_000066.1| TCCTGGTGAGGAGCTGCT

ref|NC_000023.9| TCCTGGTGAGGAGCTGCT

ref|AC_000155.1| TCCTGGTGAGGAGCTGCT

******************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS1073(REN90074)	NC_000023	153482143--153482182	20.0	100	80
	AC_000066	153988955--153988992	19.0	100	76
	AC_000155	142374152--142374191	20.0	100	80

*WARNING!*
The extragenic dinucleotide DXS1073(REN90074) locus has been reported in gametic (linkage) EQUILIBRIUM with the REN91100(F8Int13.2), F8Int22 and DXS1108 loci^[1].
^[1]		Sanchez-Garcia JF, Gallardo D, Ramirez L, Vidal F. Multiplex fluorescent analysis of four short tandem repeats for rapid haemophilia A molecular diagnosis. Thromb Haemost 2005; 94(5): 1099-103.
New!		This locus has recently been tried-and-tested as informative using the pair of primers here provided^[2].
^[2]		Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.
New!		This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[3]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.73 and 0.51, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[3]		Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

	DXS1073(REN90074)		Information
	Amplicon		132-134 bp
	Forward ampliprimer		AAGAATGCCCTCTCCGAGTT
	Reverse ampliprimer		ATTGGTGGCCTTTGAAACAC

>ref|NC_000023.9|

AAGAATGCCCTCTCCGAGTTATTACAAAGAAGCACAGAATCCCAGCCCAGTCTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGGTGGCGGGGCAAGGGGAGCAGTGTTTCAAAGGCCACCAAT

>ref|AC_000066.1|

AAGAATGCCCTCTCCGAGTTATTACAAAGAAGCACAGAATCCCAGCCCAGTCTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGGTGGCGGGGCAAGGGGAGCAGTGTTTCAAAGGCCACCAAT

>ref|AC_000155.1|

AAGAATGCCCTCTCCGAGTTATTACAAAGAAGCACAGAATCCCAGCCCAGTCTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

TGTGGTGGCGGGGCAAGGGGAGCAGTGTTTCAAAGGCCACCAAT

DXS1073(REN90074)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 134 bp

Sequence 2: ref|AC_000066.1| 132 bp

Sequence 3: ref|AC_000155.1| 134 bp

ref|NC_000023.9| AAGAATGCCCTCTCCGAGTTATTACAAAGAAGCACAGAATCCCAGCCCAGTCTCTGTGTG

ref|AC_000155.1| AAGAATGCCCTCTCCGAGTTATTACAAAGAAGCACAGAATCCCAGCCCAGTCTCTGTGTG

ref|AC_000066.1| AAGAATGCCCTCTCCGAGTTATTACAAAGAAGCACAGAATCCCAGCCCAGTCTCTGTGTG

************************************************************

ref|NC_000023.9| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGGTGGCGGGGCAAGGGGAGCAGTGTTT

ref|AC_000155.1| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGGTGGCGGGGCAAGGGGAGCAGTGTTT

ref|AC_000066.1| TGTGTGTGTGTGTGTGTGTGTGTGTGTGTGT--GGTGGCGGGGCAAGGGGAGCAGTGTTT

******************************* ***************************

ref|NC_000023.9| CAAAGGCCACCAAT

ref|AC_000155.1| CAAAGGCCACCAAT

ref|AC_000066.1| CAAAGGCCACCAAT

**************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90085	NC_000023	153484584--153484617	8.5	100	68
	AC_000066	153991394--153991427	8.5	100	68
	AC_000155	142376593--142376626	8.5	100	68

REN90085	Information
Amplicon	223 bp
Forward ampliprimer	ACCATTCCCTCCCTGACTCT
Reverse ampliprimer	GCCAGACCCCATCTCTCAAG

>ref|NC_000023.9|

ACCATTCCCTCCCTGACTCTCTCCTAACTCTGTCCCCGTCACTCCCACTCGGCGACCTGGCTCGCCCTTTGCCCTCTGCTCTTCTCACTC

TATGCCTGACCCTGCACTAACTGTAGAAAATGCAGAAATGAGAAACAGACATTCATTCAACAAGCATTTATTCTTATTTATTTATTTATT

TATTTATTTATTTATTTTTATTTCTTGAGAGATGGGGTCTGGC

>ref|AC_000066.1|

ACCATTCCCTCCCTGACTCTCTCCTAACTCTGTCCCCGTCACTCCCACTCGGCGACCTGGCTCGCCCTTTGCCCTCTGCTCTTCTCACTC

TATGCCTGACCCTGCACTAACTGTAGAAAATGCAGAAATGAGAAACAGACATTCATTCAACAAGCATTTATTCTTATTTATTTATTTATT

TATTTATTTATTTATTTTTATTTCTTGAGAGATGGGGTCTGGC

>ref|AC_000155.1|

ACCATTCCCTCCCTGACTCTCTCCTAACTCTGTCCCCGTCACTCCCACTCGGCGACCTGGCTCGCCCTTTGCCCTCTGCTCTTCTCACTC

TATGCCTGACCCTGCACTAACTGTAGAAAATGCAGAAATGAGAAACAGACATTCATTCAACAAGCATTTATTCTTATTTATTTATTTATT

TATTTATTTATTTATTTTTATTTCTTGAGAGATGGGGTCTGGC

REN90085

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 223 bp

Sequence 2: ref|AC_000066.1| 223 bp

Sequence 3: ref|AC_000155.1| 223 bp

ref|NC_000023.9| ACCATTCCCTCCCTGACTCTCTCCTAACTCTGTCCCCGTCACTCCCACTCGGCGACCTGG

ref|AC_000155.1| ACCATTCCCTCCCTGACTCTCTCCTAACTCTGTCCCCGTCACTCCCACTCGGCGACCTGG

ref|AC_000066.1| ACCATTCCCTCCCTGACTCTCTCCTAACTCTGTCCCCGTCACTCCCACTCGGCGACCTGG

************************************************************

ref|NC_000023.9| CTCGCCCTTTGCCCTCTGCTCTTCTCACTCTATGCCTGACCCTGCACTAACTGTAGAAAA

ref|AC_000155.1| CTCGCCCTTTGCCCTCTGCTCTTCTCACTCTATGCCTGACCCTGCACTAACTGTAGAAAA

ref|AC_000066.1| CTCGCCCTTTGCCCTCTGCTCTTCTCACTCTATGCCTGACCCTGCACTAACTGTAGAAAA

************************************************************

ref|NC_000023.9| TGCAGAAATGAGAAACAGACATTCATTCAACAAGCATTTATTCTTATTTATTTATTTATT

ref|AC_000155.1| TGCAGAAATGAGAAACAGACATTCATTCAACAAGCATTTATTCTTATTTATTTATTTATT

ref|AC_000066.1| TGCAGAAATGAGAAACAGACATTCATTCAACAAGCATTTATTCTTATTTATTTATTTATT

************************************************************

ref|NC_000023.9| TATTTATTTATTTATTTTTATTTCTTGAGAGATGGGGTCTGGC

ref|AC_000155.1| TATTTATTTATTTATTTTTATTTCTTGAGAGATGGGGTCTGGC

ref|AC_000066.1| TATTTATTTATTTATTTTTATTTCTTGAGAGATGGGGTCTGGC

*******************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90200(HA472)	NC_000023	153553700--153553902	67.0	94	343
	AC_000066	154058587--154058776	63.0	91	292
	AC_000155	142444622--142444812	63.0	93	319

New!

This locus has recently been tried-and-tested as informative using the pair of primers here provided^[1].

^[1]

Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.

REN90200(HA472)	Information
Amplicon	256-269 bp
Forward ampliprimer	GCTCCTTTGATTGGATAATTTCA
Reverse ampliprimer	TGCCTCAACATCAGAATAGACC

>ref|NC_000023.9|

GCTCCTTTGATTGGATAATTTCAAATGTCCTGTCATTGAGCTCCCTTCTTCTTCTTTCTTCCTCTTCTTCTTCTTCTTCTTCTTCTTCTT

CTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCCTCCCCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

CTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTTCTTCTTCTTCTTCTTCATCTTCTTCTTCTTGGTCTATTCTGATGTTGAGGCA

>ref|AC_000066.1|

GCTCCTTTGATTGGATAATTTCAAATGTCCTGTCATTGAGCTCCCTTCTTCTTCTTTCTTCCTCTTCTTCTTCTTCTTCTTCTTCTTCTT

CTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCCTCCCCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

CTTCTTCCTCTGCTTCTTCTTCTTTCTTCTTCTTCTTCTCATCTTCTTCTTCTTGGTCTATTCTGATGTTGAGGCA

>ref|AC_000155.1|

GCTCCTTTGATTGGATAATTTCAAATGTCCTGTCATTGAGCTCCCTTCTTCTTCTTTCTTCCTCTTCTTCTTCTTCTTCTTCTTCTTCTT

CTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCCTCCCCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

CTTCTTCTTCTTCTTCTTCTTCTTTCTTCTTCTTCTTCTTCATCTTCTTCTTCTTGGTCTATTCTGATGTTGAGGCA

REN90200(HA472)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 269 bp

Sequence 2: ref|AC_000066.1| 256 bp

Sequence 3: ref|AC_000155.1| 257 bp

ref|AC_000066.1| GCTCCTTTGATTGGATAATTTCAAATGTCCTGTCATTGAGCTCCCTTCTTCTTCTTTCTT

ref|AC_000155.1| GCTCCTTTGATTGGATAATTTCAAATGTCCTGTCATTGAGCTCCCTTCTTCTTCTTTCTT

ref|NC_000023.9| GCTCCTTTGATTGGATAATTTCAAATGTCCTGTCATTGAGCTCCCTTCTTCTTCTTTCTT

************************************************************

ref|AC_000066.1| CCTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

ref|AC_000155.1| CCTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

ref|NC_000023.9| CCTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

************************************************************

ref|AC_000066.1| CTTCTTCTTCTTCTTC------------TTCTTCCTCCCCTTCTTCTTCTTCTTCTTCTT

ref|AC_000155.1| CTTCTTCTTCTTCTTC------------TTCTTCCTCCCCTTCTTCTTCTTCTTCTTCTT

ref|NC_000023.9| CTTCTTCTTCTTCTTCCTCCCCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTT

**************** ****** ** *********************

ref|AC_000066.1| CTTCTTCTTCTTCTTCTTCCTCTGCTTCTTCTTCTTTCTTCTTCTTCTTCT-CATCTTCT

ref|AC_000155.1| CTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTTCTTCTTCTTCTTCTTCATCTTCT

ref|NC_000023.9| CTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTTCTTCTTCTTCTTCTTCATCTTCT

******************* *** *************************** ********

ref|AC_000066.1| TCTTCTTGGTCTATTCTGATGTTGAGGCA

ref|AC_000155.1| TCTTCTTGGTCTATTCTGATGTTGAGGCA

ref|NC_000023.9| TCTTCTTGGTCTATTCTGATGTTGAGGCA

*****************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604240/REN90371(GAB3Int2.2)	NC_000023	153595212--153595255	22.0	100	88
	AC_000066	154100601--154100644	22.0	100	88
	AC_000155	142485953--142485988	18.0	100	72

New!

WARNING!

This microsatellite marker was firstly predicted In-Silico as informative ^[1], and it has recently been tried-and-tested, apparently (no primer sequences provided), using a pair set of ampliprimers different to the one here provided ^[2]. The authors reported that genotyping of this STR marker yielded peak patterns that made allele calling difficult and, therefore, excluded it form the multiplex assay. Worth of note, the latter study also reported similar impairing effects for two other STR marker loci (stSG604469 and REN89898), all of which contain a [AT] repeat unit.

^[1]

^[2]

stSG604240/REN90371(GAB3Int2.2)	Information
Amplicon	219-227 bp
Forward ampliprimer	CTTCTTTATGGCCCTCAGCA
Reverse ampliprimer	TCTGGGCACATGGTAGATGTAA

>ref|NC_000023.9|

CTTCTTTATGGCCCTCAGCAGCAGCCTTGCCTCATTGTTTTTTCACTCAACTAACAAACATATATATATATATATATATATATATATATA

TATATATATATATGCATAACAAGTACATGCATATAGTTATATATGTACTTGTGCATATATGTATGTATACCTATACATGTTTATTATATA

GGTATATGTCTATATAGAATACCTATTACATCTACCATGTGCCCAGA

>ref|AC_000066.1|

CTTCTTTATGGCCCTCAGCAGCAGCCTTGCCTCATTGTTTTTTCACTCAACTAACAAACATATATATATATATATATATATATATATATA

TATATATATATATGCATAACAAGTACATGCATATAGTTATATATGTACTTGTGCATATATGTATGTATACCTATACATGTTTATTATATA

GGTATATGTCTATATAGAATACCTATTACATCTACCATGTGCCCAGA

>ref|AC_000155.1|

CTTCTTTATGGCCCTCAGCAGCAGCCTTGCCTCATTGTTTTTTCACTCAACTAACAAACATATATATATATATATATATATATATATATA

TATATGCATAACAAGTACATGCATATAGTTATATATGTACTTGTGCATATATGTATGTATACCTATACATGTTTATTATATAGGTATATG

TCTATATAGAATACCTATTACATCTACCATGTGCCCAGA

stSG604240/REN90371(GAB3Int2)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 227 bp

Sequence 2: ref|AC_000066.1| 227 bp

Sequence 3: ref|AC_000155.1| 219 bp

ref|NC_000023.9| CTTCTTTATGGCCCTCAGCAGCAGCCTTGCCTCATTGTTTTTTCACTCAACTAACAAACA

ref|AC_000066.1| CTTCTTTATGGCCCTCAGCAGCAGCCTTGCCTCATTGTTTTTTCACTCAACTAACAAACA

ref|AC_000155.1| CTTCTTTATGGCCCTCAGCAGCAGCCTTGCCTCATTGTTTTTTCACTCAACTAACAAACA

************************************************************

ref|NC_000023.9| TATATATATATATATATATATATATATATATATATATATATATGCATAACAAGTACATGC

ref|AC_000066.1| TATATATATATATATATATATATATATATATATATATATATATGCATAACAAGTACATGC

ref|AC_000155.1| TATATATATATATATATATATATATATATA--------TATATGCATAACAAGTACATGC

****************************** **********************

ref|NC_000023.9| ATATAGTTATATATGTACTTGTGCATATATGTATGTATACCTATACATGTTTATTATATA

ref|AC_000066.1| ATATAGTTATATATGTACTTGTGCATATATGTATGTATACCTATACATGTTTATTATATA

ref|AC_000155.1| ATATAGTTATATATGTACTTGTGCATATATGTATGTATACCTATACATGTTTATTATATA

************************************************************

ref|NC_000023.9| GGTATATGTCTATATAGAATACCTATTACATCTACCATGTGCCCAGA

ref|AC_000066.1| GGTATATGTCTATATAGAATACCTATTACATCTACCATGTGCCCAGA

ref|AC_000155.1| GGTATATGTCTATATAGAATACCTATTACATCTACCATGTGCCCAGA

***********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604242/REN90378(GAB3Int2.1)	NC_000023	153596862--153596895	17.0	81	41
	NC_000023	153596898--153596924	13.5	100	54
	AC_000066	153628186--154102284	17.0	81	41
	AC_000066	154102287--154102313	13.5	100	54
	AC_000155	142044301--142487628	17.0	81	41
	AC_000155	142487631--142487657	13.5	100	54

*stSG604242/REN90378(GAB3Int2.1)*	Information
Amplicon	215 bp
Forward ampliprimer	AATGTCAGGGGCTCAACAAC
Reverse ampliprimer	TAAGTTTGCTCTGGGCCATT

>ref|NC_000023.9|

AATGTCAGGGGCTCAACAACAAAATCCATGAAGGGAAGTTAATTGTAACCCTAGGTTACAATATACCTGGAAAATTACATAGGATGAGTT

GTGTGTGTGTGCGTGTGTGCGTGTGTGCGTGTGGCAGAGAGAGAGAGAGAGAGAGAGAGAGACTCTGGGGTACTCTTCATGGGTCAATTG

CTCCCACTGCCCAGAAATGGCCCAGAGCAAACTTA

>ref|AC_000066.1|

AATGTCAGGGGCTCAACAACAAAATCCATGAAGGGAAGTTAATTGTAACCCTAGGTTACAATATACCTGGAAAATTACATAGGATGAGTT

GTGTGTGTGTGCGTGTGTGCGTGTGTGCGTGTGGCAGAGAGAGAGAGAGAGAGAGAGAGAGACTCTGGGGTACTCTTCATGGGTCAATTG

CTCCCACTGCCCAGAAATGGCCCAGAGCAAACTTA

>ref|AC_000155.1|

AATGTCAGGGGCTCAACAACAAAATCCATGAAGGGAAGTTAATTGTAACCCTAGGTTACAATATACCTGGAAAATTACATAGGATGAGTT

GTGTGTGTGTGCGTGTGTGCGTGTGTGCGTGTGGCAGAGAGAGAGAGAGAGAGAGAGAGAGACTCTGGGGTACTCTTCATGGGTCAATTG

CTCCCACTGCCCAGAAATGGCCCAGAGCAAACTTA

stSG604242/REN90378(GAB3Int2.1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 215 bp

Sequence 2: ref|AC_000066.1| 215 bp

Sequence 3: ref|AC_000155.1| 215 bp

ref|NC_000023.9| AATGTCAGGGGCTCAACAACAAAATCCATGAAGGGAAGTTAATTGTAACCCTAGGTTACA

ref|AC_000155.1| AATGTCAGGGGCTCAACAACAAAATCCATGAAGGGAAGTTAATTGTAACCCTAGGTTACA

ref|AC_000066.1| AATGTCAGGGGCTCAACAACAAAATCCATGAAGGGAAGTTAATTGTAACCCTAGGTTACA

************************************************************

ref|NC_000023.9| ATATACCTGGAAAATTACATAGGATGAGTTGTGTGTGTGTGCGTGTGTGCGTGTGTGCGT

ref|AC_000155.1| ATATACCTGGAAAATTACATAGGATGAGTTGTGTGTGTGTGCGTGTGTGCGTGTGTGCGT

ref|AC_000066.1| ATATACCTGGAAAATTACATAGGATGAGTTGTGTGTGTGTGCGTGTGTGCGTGTGTGCGT

************************************************************

ref|NC_000023.9| GTGGCAGAGAGAGAGAGAGAGAGAGAGAGAGACTCTGGGGTACTCTTCATGGGTCAATTG

ref|AC_000155.1| GTGGCAGAGAGAGAGAGAGAGAGAGAGAGAGACTCTGGGGTACTCTTCATGGGTCAATTG

ref|AC_000066.1| GTGGCAGAGAGAGAGAGAGAGAGAGAGAGAGACTCTGGGGTACTCTTCATGGGTCAATTG

************************************************************

ref|NC_000023.9| CTCCCACTGCCCAGAAATGGCCCAGAGCAAACTTA

ref|AC_000155.1| CTCCCACTGCCCAGAAATGGCCCAGAGCAAACTTA

ref|AC_000066.1| CTCCCACTGCCCAGAAATGGCCCAGAGCAAACTTA

***********************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604248/REN90414(GAB3Int1)	NC_000023	153605812--153605855	11.0	100	88
	AC_000066	154111201--154111244	11.0	100	88
	AC_000155	142496574--142496617	11.0	100	88

New!	This microsatellite marker has been recently validated experimentally as informative, albeit with low heterozygosity rate ^[1].
New!	Multiallelic interlocus non-random association analysis revealed that **GAB3Int1 is at gametic (linkage) EQUILIBRIUM with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1*.1. The observed breakdown of gametic disequilibrium attests historical recombination between GAB3*Int1 and the F8 gene.
^[1]	Machado FB, Alves Da Silva AF, Rossetti LC, De Brasi CD, Medina-Acosta E. Informativeness of a novel multiallelic marker-set comprising an F8 intron 21 and three tightly linked loci for haemophilia A carriership analysis. Haemophilia 2011, 17:257-66.

*stSG604248/REN90414(GAB3Int1)*	Information
Amplicon	147 bp
Forward ampliprimer	CCTGGGCAACAGAGTGAGAC
Reverse ampliprimer	GAAGGGGAGAAGCAGTTTCC

>ref|NC_000023.9|

CCTGGGCAACAGAGTGAGACTTCATCTTAAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAAAATAAAATGACAATGTGAC

TGTTGGCCCCCAGGGCTCATGGCATCACTTGCCTTGGGGAAACTGCTTCTCCCCTTC

>ref|AC_000066.1|

CCTGGGCAACAGAGTGAGACTTCATCTTAAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAAAATAAAATGACAATGTGAC

TGTTGGCCCCCAGGGCTCATGGCATCACTTGCCTTGGGGAAACTGCTTCTCCCCTTC

>ref|AC_000155.1|

CCTGGGCAACAGAGTGAGACTTCATCTTAAATAAATAAATAAATAAATAAATAAATAAATAAATAAATAAAAATAAAATGACAATGTGAC

TGTTGGCCCCCAGGGCTCATGGCATCACTTGCCTTGGGGAAACTGCTTCTCCCCTTC

stSG604248/REN90414(GAB3Int1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 147 bp

Sequence 2: ref|AC_000066.1| 147 bp

Sequence 3: ref|AC_000155.1| 147 bp

ref|NC_000023.9| CCTGGGCAACAGAGTGAGACTTCATCTTAAATAAATAAATAAATAAATAAATAAATAAAT

ref|AC_000155.1| CCTGGGCAACAGAGTGAGACTTCATCTTAAATAAATAAATAAATAAATAAATAAATAAAT

ref|AC_000066.1| CCTGGGCAACAGAGTGAGACTTCATCTTAAATAAATAAATAAATAAATAAATAAATAAAT

************************************************************

ref|NC_000023.9| AAATAAATAAAAATAAAATGACAATGTGACTGTTGGCCCCCAGGGCTCATGGCATCACTT

ref|AC_000155.1| AAATAAATAAAAATAAAATGACAATGTGACTGTTGGCCCCCAGGGCTCATGGCATCACTT

ref|AC_000066.1| AAATAAATAAAAATAAAATGACAATGTGACTGTTGGCCCCCAGGGCTCATGGCATCACTT

************************************************************

ref|NC_000023.9| GCCTTGGGGAAACTGCTTCTCCCCTTC

ref|AC_000155.1| GCCTTGGGGAAACTGCTTCTCCCCTTC

ref|AC_000066.1| GCCTTGGGGAAACTGCTTCTCCCCTTC

***************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604294/REN90682/UniSTS:68640/HSC269_(M1) (MPP1Int5)	NC_000023	153668571--153668616	23.0	100	92
	AC_000066	154173942--154173979	19.0	100	76

New!	This microsatellite marker was firstly predicted In-Silico as informative ^[1], and recently validated experimentally under a different coined name by Ding et al. (2012) ^[2]. *WARNING!* We recently warrant about the need to acknowledge accurate tracking genetic markers in haemophilia linkage analysis to avoid unnecessary priority claims, as in this case ^[3].
^[1]	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.
^[2]	Ding QL, Lu YL, Dai J, Xi XD, Wan XF, Wang HL. Characterisation and validation of a novel panel of the six short tandem repeats for genetic counselling in Chinese haemophilia A pedigrees. Haemophilia 2012, 18(4):621-625
^[3]	Machado FB, Medina-Acosta E (2012) Accurate tracking genetic markers in haemophilia linkage analysis: a matter beyond scientific priority. Haemophilia 18:e359--e360
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[4]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.68 and 0.53, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[4]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

*stSG604294/REN90682/UniSTS:68640/HSC269_(M1) (MPP1Int5)*	Information
Amplicon	273-281 bp
Forward ampliprimer	GGATCTGGGTCGTCTGTCTC
Reverse ampliprimer	GAGTTGGGAAGTGCCATCAT

>ref|NC_000023.9|

GGATCTGGGTCGTCTGTCTCAGTACAGGGCAGCTACTCTTTGAACAACTGTTTTCACAATTGAAAATATGGACACTCATATCTATCAAAG

AACTTGAATTTATTATTGAAAATTCTCCTCTCCTGCTCCCAAACACACACACACACACACACACACACACACACACACACACACACACCT

ACCTCATTACTTCATTGACAAATTTTCCAAGCATCATTTGAAGAAAGAAGGCTAATCTCATACGAAAACAGATTATAGAAAATGATGGCA

CTTCCCAACTC

>ref|AC_000066.1|

GGATCTGGGTCGTCTGTCTCAGTACAGGGCAGCTACTCTTTGAACAACTGTTTTCACAATTGAAAATATGGACACTCATATCTATCAAAG

AACTTGAATTTATTATTGAAAATTCTCCTCTCCTGCTCCCAAACACACACACACACACACACACACACACACACACACACCTACCTCATT

ACTTCATTGACAAATTTTCCAAGCATCATTTGAAGAAAGAAGGCTAATCTCATACGAAAACAGATTATAGGAAATGATGGCACTTCCCAA

CTC

stSG604294/REN90682/UniSTS:68640/HSC269_(M1) (MPP1Int5)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 281 bp

Sequence 2: ref|AC_000066.1| 273 bp

ref|NC_000023.9| GGATCTGGGTCGTCTGTCTCAGTACAGGGCAGCTACTCTTTGAACAACTGTTTTCACAAT

ref|AC_000066.1| GGATCTGGGTCGTCTGTCTCAGTACAGGGCAGCTACTCTTTGAACAACTGTTTTCACAAT

************************************************************

ref|NC_000023.9| TGAAAATATGGACACTCATATCTATCAAAGAACTTGAATTTATTATTGAAAATTCTCCTC

ref|AC_000066.1| TGAAAATATGGACACTCATATCTATCAAAGAACTTGAATTTATTATTGAAAATTCTCCTC

************************************************************

ref|NC_000023.9| TCCTGCTCCCAAACACACACACACACACACACACACACACACACACACACACACACACCT

ref|AC_000066.1| TCCTGCTCCCAAACACACACACACACACACACACACACACACACACA--------CACCT

*********************************************** *****

ref|NC_000023.9| ACCTCATTACTTCATTGACAAATTTTCCAAGCATCATTTGAAGAAAGAAGGCTAATCTCA

ref|AC_000066.1| ACCTCATTACTTCATTGACAAATTTTCCAAGCATCATTTGAAGAAAGAAGGCTAATCTCA

************************************************************

ref|NC_000023.9| TACGAAAACAGATTATAGAAAATGATGGCACTTCCCAACTC

ref|AC_000066.1| TACGAAAACAGATTATAGGAAATGATGGCACTTCCCAACTC

****************** **********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90833	NC_000023	153694353--153694393	10.3	100	82
	AC_000066	154199716--154199744	7.3	100	58
	AC_000155	142584435--142584463	7.3	100	58

New!	This microsatellite marker has been recently validated experimentally as informative,^[1] albeit using a different (redesigned) pair of ampliprimers. *WARNING!* The redesigned REN90833 ampliprimers also anneal (reverse primer with 3bp 5’ mismatched to a non-STR locus on chromosome 10, yielding a 106 bp non-polymorphic PCR product with DNA from both males and females). This product does not interfere with the designation of REN90833 ChrX alleles, which ranged 138-160 bp in the population subset tested.
New!	Extended haplotype homozygosity (HEE) analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[2]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect).
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[4]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.49 and 0.43, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.
^[2]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.
^[3]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

REN90833	Information
Amplicon	118-130 bp
Forward ampliprimer	GCAAATTGTGAGGGAGGTATG
Reverse ampliprimer	GAACTGCATTGGGCAAATCT

REN90833 (redesigned) ^[1]	Information
Amplicon	138-160 bp + 106bp non-polymorphic product
Forward ampliprimer	GGCAAATTGTGAGGGAGGTA
Reverse ampliprimer	CCAAAGGGAAAAGTCAAGCA

>ref|NC_000023.9|

GCAAATTGTGAGGGAGGTATGTAGCTTACTTATTTATTTATTTATTTATTTATTTATTTATTTATTTATTAATCTTTATAGCTATCTTA

TTTAGGAATAGAATGGGAGGGAGATTTGCCCAATGCAGTTC

>ref|AC_000066.1|

GCAAATTGTGAGGGAGGTATGTAGCTTACTTATTTATTTATTTATTTATTTATTTATTAATCTTTATAGCTATCTTATTTAGGAATAGA

ATGGGAGGGAGATTTGCCCAATGCAGTTC

>ref|AC_000155.1|

GCAAATTGTGAGGGAGGTATGTAGCTTACTTATTTATTTATTTATTTATTTATTTATTAATCTTTATAGCTATCTTATTTAGGAATAGA

ATGGGAGGGAGATTTGCCCAATGCAGTTC

REN90833 (redesigned) ^[1]validation by iPCR at UCSC

>chrX:153604833+153604985 153bp GGCAAATTGTGAGGGAGGTA CCAAAGGGAAAAGTCAAGCA

GGCAAATTGTGAGGGAGGTAtgtagcttacttatttatttatttatttat

ttatttatttatttatttattaatctttatagctatcttatttaggaata

gaatgggagggagatttgcccaatgcagttcccTGCTTGACTTTTCCCTT

TGG

Note:

Monomorphic 106 bp mapping at Chr10 (reverse primer with 3bp 5’ mismatched to a non-STR locus on chromosome 10, yielding a 106 bp non-polymorphic PCR product with DNA from both males and females)

>chr10:96372691+96372796 106bp GGCAAATTGTGAGGGAGGTA CCAAAGGGAAAAGTCAAGCA

GGCAAATTGTGAGGGAGGTAtgtagctcttttatctttttagctatctta

ttttggaataaaatgggaggcaggtttgtagttcccTGCTTGACTTTTCC

CTTgac

REN90833

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 130 bp

Sequence 2: ref|AC_000066.1| 118 bp

Sequence 3: ref|AC_000155.1| 118 bp

ref|AC_000066.1| GCAAATTGTGAGGGAGGTATGTAGCTTACTTATTTATTTATTTATTTATTTATTTATT--

ref|AC_000155.1| GCAAATTGTGAGGGAGGTATGTAGCTTACTTATTTATTTATTTATTTATTTATTTATT--

ref|NC_000023.9| GCAAATTGTGAGGGAGGTATGTAGCTTACTTATTTATTTATTTATTTATTTATTTATTTA

**********************************************************

ref|AC_000066.1| ----------AATCTTTATAGCTATCTTATTTAGGAATAGAATGGGAGGGAGATTTGCCC

ref|AC_000155.1| ----------AATCTTTATAGCTATCTTATTTAGGAATAGAATGGGAGGGAGATTTGCCC

ref|NC_000023.9| TTTATTTATTAATCTTTATAGCTATCTTATTTAGGAATAGAATGGGAGGGAGATTTGCCC

**************************************************

ref|AC_000066.1| AATGCAGTTC

ref|AC_000155.1| AATGCAGTTC

ref|NC_000023.9| AATGCAGTTC

**********

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90932(F8Int25.3)	NC_000023	153731553--153731589	18.5	100	74
	AC_000066	154236906--154236942	18.5	100	74
	AC_000155	142622013--142622049	18.5	100	74

New!

This locus has recently been tried-and-tested as informative using the pair of primers here provided^[1].

^[1]

Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.

REN90932(F8Int25.3)	Information
Amplicon	122 bp
Forward ampliprimer	TCCAAGATCAAGGGGTAGGC
Reverse ampliprimer	GCCTGGACTACAGAGGGAGA

>ref|NC_000023.9|

TCCAAGATCAAGGGGTAGGCAGGCTTGTTTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATTTTTTTTTTGTCTGTTTGTTT

TTTGAGATGGAGTCTCCCTCTGTAGTCCAGGC

>ref|AC_000066.1|

TCCAAGATCAAGGGGTAGGCAGGCTTGTTTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATTTTTTTTTTGTCTGTTTGTTT

TTTGAGATGGAGTCTCCCTCTGTAGTCCAGGC

>ref|AC_000155.1|

TCCAAGATCAAGGGGTAGGCAGGCTTGTTTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATTTTTTTTTTGTCTGTTTGTTT

TTTGAGATGGAGTCTCCCTCTGTAGTCCAGGC

REN90932(F8Int25.3)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 122 bp

Sequence 2: ref|AC_000066.1| 122 bp

Sequence 3: ref|AC_000155.1| 122 bp

ref|AC_000066.1| TCCAAGATCAAGGGGTAGGCAGGCTTGTTTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|NC_000023.9| TCCAAGATCAAGGGGTAGGCAGGCTTGTTTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

ref|AC_000155.1| TCCAAGATCAAGGGGTAGGCAGGCTTGTTTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG

************************************************************

ref|AC_000066.1| TGTGTGTATTTTTTTTTTGTCTGTTTGTTTTTTGAGATGGAGTCTCCCTCTGTAGTCCAG

ref|NC_000023.9| TGTGTGTATTTTTTTTTTGTCTGTTTGTTTTTTGAGATGGAGTCTCCCTCTGTAGTCCAG

ref|AC_000155.1| TGTGTGTATTTTTTTTTTGTCTGTTTGTTTTTTGAGATGGAGTCTCCCTCTGTAGTCCAG

************************************************************

ref|AC_000066.1| GC

ref|NC_000023.9| GC

ref|AC_000155.1| GC

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604314/REN90841(F8Int25.2)	NC_000023	153736060--153736088	14.5	100	58
	AC_000066	154241413--154241441	14.5	100	58
	AC_000155	142626521--142626553	16.5	100	66

*WARNING!*
This microsatellite marker has been recently validated experimentally as informative ^[1,2]. More recently, it was genotyped under a different coined name by Ding et al. (2012) ^[3]. We recently warned about the need to acknowledge accurate tracking genetic markers in haemophilia linkage analysis to avoid unnecessary priority claims, as in this case ^[4].
New!	Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[5]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect).
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[6]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.5 and 0.47, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Duarte LP, Medina-Acosta E. A novel informative dinucleotide microsatellite marker located on human factor VIII intron 25. Haemophilia 2009, 15:613-614.
^[2]	de Sá Machado Araújo G, Louvain de Souza T, Medina-Acosta E. Use of SNPSTR combined markers for gametic phase determination of X chromosomes in Hemophilia A female carriers. In 58o. Congresso Brasileiro de Genética. 2012. Foz do Iguaçu: Sociedade Brasileira de Genética.
^[3]	Ding QL, Lu YL, Dai J, Xi XD, Wan XF, Wang HL. Characterisation and validation of a novel panel of the six short tandem repeats for genetic counselling in Chinese haemophilia A pedigrees. Haemophilia 2012, 18(4):621-625
^[4]	Machado FB, Medina-Acosta E (2012) Accurate tracking genetic markers in haemophilia linkage analysis: a matter beyond scientific priority. Haemophilia 18:e359--e360
^[5]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.
^[6]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

stSG604314/REN90841(F8Int25.2)	Information
Amplicon	144-148 bp
Forward ampliprimer	CTCTAGGCTGCCAACTCACC
Reverse ampliprimer	TCATGGTTCCCAAAGAAACA

>ref|NC_000023.9|

CTCTAGGCTGCCAACTCACCCTGCAGATCTTGAGACTTGGCAGACTCCATAATCACATGAGCTAATTCCTTGTGTGTGTGTGTGTGTGTG

TGTGTGTGTATGTATCTCCTCTTGGTTCTGGTTTTGTTTCTTTGGGAACCATGA

>ref|AC_000066.1|

CTCTAGGCTGCCAACTCACCCTGCAGATCTTGAGACTTGGCAGACTCCATAATCACATGAGCTAATTCCTTGTGTGTGTGTGTGTGTGTG

TGTGTGTGTATGTATCTCCTCTTGGTTCTGGTTTTGTTTCTTTGGGAACCATGA

>ref|AC_000155.1|

CTCTAGGCTGCCAACTCACCCTGCAGATCTTGAGACTTGGCAGACTCCATAATCACATGAGCTAATTCCTTGTGTGTGTGTGTGTGTGTG

TGTGTGTGTGTGTATGTATCTCCTCTTGGTTCTGGTTTTGTTTCTTTGGGAACCATGA

stSG604314/REN90841(F8Int25.2)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 144 bp

Sequence 2: ref|AC_000066.1| 144 bp

Sequence 3: ref|AC_000155.1| 148 bp

ref|NC_000023.9| CTCTAGGCTGCCAACTCACCCTGCAGATCTTGAGACTTGGCAGACTCCATAATCACATGA

ref|AC_000066.1| CTCTAGGCTGCCAACTCACCCTGCAGATCTTGAGACTTGGCAGACTCCATAATCACATGA

ref|AC_000155.1| CTCTAGGCTGCCAACTCACCCTGCAGATCTTGAGACTTGGCAGACTCCATAATCACATGA

************************************************************

ref|NC_000023.9| GCTAATTCCTTGTGTGTGTGTGTGTGTGTGTGTGTGTGT----ATGTATCTCCTCTTGGT

ref|AC_000066.1| GCTAATTCCTTGTGTGTGTGTGTGTGTGTGTGTGTGTGT----ATGTATCTCCTCTTGGT

ref|AC_000155.1| GCTAATTCCTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATGTATCTCCTCTTGGT

*************************************** *****************

ref|NC_000023.9| TCTGGTTTTGTTTCTTTGGGAACCATGA

ref|AC_000066.1| TCTGGTTTTGTTTCTTTGGGAACCATGA

ref|AC_000155.1| TCTGGTTTTGTTTCTTTGGGAACCATGA

****************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90957(F8Int25.1)	NC_000023	153740631--153740651	10.5	89	33
	AC_000066	154245986--154246006	10.5	89	33
	AC_000155	142631097--142631117	10.5	89	33

New!
This microsatellite marker has been recently validated experimentally as uninformative,^[1] and therefore not suitable for linkage analysis.
^[1]	de Sá Machado Araújo G, Louvain de Souza T, Medina-Acosta E. Use of SNPSTR combined markers for gametic phase determination of X chromosomes in Hemophilia A female carriers. In 58o. Congresso Brasileiro de Genética. 2012. Foz do Iguaçu: Sociedade Brasileira de Genética.

REN90957(F8Int25.1)	Information
Amplicon	145 bp
Forward ampliprimer	CTGCCTTTGGACTGGAACAT
Reverse ampliprimer	TGTGTTAGTCAGGGATATCCAAA

>ref|NC_000023.9|

CTGCCTTTGGACTGGAACATCAGCTTTTTCCTGCTTTCAGTCTTGAACTAAAACAGTTTAGCTCATCCAGAGAGACAGAGAGAGAGAGAC

AGACAGACAGAATCTCCTATTGGTTCTATTTCTTTGGATATCCCTGACTAACACA

>ref|AC_000066.1|

CTGCCTTTGGACTGGAACATCAGCTTTTTCCTGCTTTCAGTCTTGAACTAAAACAGTTTAGCTCATCCAGAGAGACAGAGAGAGAGAGAC

AGACAGACAGAATCTCCTATTGGTTCTATTTCTTTGGATATCCCTGACTAACACA

>ref|AC_000155.1|

CTGCCTTTGGACTGGAACATCAGCTTTTTCCTGCTTTCAGTCTTGAACTAAAACAGTTTAGCTCATCCAGAGAGACAGAGAGAGAGAGAC

AGACAGACAGAATCTCCTATTGGTTCTATTTCTTTGGATATCCCTGACTAACACA

REN90957(F8Int25.1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 145 bp

Sequence 2: ref|AC_000066.1| 145 bp

Sequence 3: ref|AC_000155.1| 145 bp

ref|NC_000023.9| CTGCCTTTGGACTGGAACATCAGCTTTTTCCTGCTTTCAGTCTTGAACTAAAACAGTTTA

ref|AC_000155.1| CTGCCTTTGGACTGGAACATCAGCTTTTTCCTGCTTTCAGTCTTGAACTAAAACAGTTTA

ref|AC_000066.1| CTGCCTTTGGACTGGAACATCAGCTTTTTCCTGCTTTCAGTCTTGAACTAAAACAGTTTA

************************************************************

ref|NC_000023.9| GCTCATCCAGAGAGACAGAGAGAGAGAGACAGACAGACAGAATCTCCTATTGGTTCTATT

ref|AC_000155.1| GCTCATCCAGAGAGACAGAGAGAGAGAGACAGACAGACAGAATCTCCTATTGGTTCTATT

ref|AC_000066.1| GCTCATCCAGAGAGACAGAGAGAGAGAGACAGACAGACAGAATCTCCTATTGGTTCTATT

************************************************************

ref|NC_000023.9| TCTTTGGATATCCCTGACTAACACA

ref|AC_000155.1| TCTTTGGATATCCCTGACTAACACA

ref|AC_000066.1| TCTTTGGATATCCCTGACTAACACA

*************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN90965(F8Int24.1)	NC_000023	153742466--153742512	12.3	82	44
	NC_000023	153742517--153742535	9.5	100	38
	AC_000066	154247821--154247867	12.3	82	44
	AC_000066	154247872--154247890	9.5	100	38
	AC_000155	142632932--142632978	12.3	82	44
	AC_000155	142632983--142633001	9.5	100	38

New!
This microsatellite marker has been recently validated experimentally as uninformative,^[1] and therefore not suitable for linkage analysis.
^[1]	de Sá Machado Araújo G, Louvain de Souza T, Medina-Acosta E. Use of SNPSTR combined markers for gametic phase determination of X chromosomes in Hemophilia A female carriers. In 58o. Congresso Brasileiro de Genética. 2012. Foz do Iguaçu: Sociedade Brasileira de Genética.

REN90965(F8Int24.1)	Information
Amplicon	287 bp
Forward ampliprimer	ATGAGGTTTCACCAGGTTGG
Reverse ampliprimer	GGCAGGATAATCGCATGAAC

>ref|NC_000023.9|

ATGAGGTTTCACCAGGTTGGCCAGGCTGGTCTTGAACTCCTGACTTCAGGTGATCTGCCACCTAGGCCTCCCAAAGTGCTAGGATTACAG

GCGTGAGCCACTCTGCCCGGCCTCCTTCCTTCCTTCCTTTCTTTCTTTTTCTTCCTTCCTTCTTTCCTTTCTTTCTCTCTCTCTCTCTCT

CTGCCTCTTTCAGACAGAGTCAGGCACTGTTGCCTGGGCTGGAGTGCAATAGCATGATCTCAGCTCACTGCAACCTCCGCCTCCCAGGTT

CATGCGATTATCCTGCC

>ref|AC_000066.1|

ATGAGGTTTCACCAGGTTGGCCAGGCTGGTCTTGAACTCCTGACTTCAGGTGATCTGCCACCTAGGCCTCCCAAAGTGCTAGGATTACAG

GCGTGAGCCACTCTGCCCGGCCTCCTTCCTTCCTTCCTTTCTTTCTTTTTCTTCCTTCCTTCTTTCCTTTCTTTCTCTCTCTCTCTCTCT

CTGCCTCTTTCAGACAGAGTCAGGCACTGTTGCCTGGGCTGGAGTGCAATAGCATGATCTCAGCTCACTGCAACCTCCGCCTCCCAGGTT

CATGCGATTATCCTGCC

>ref|AC_000155.1|

ATGAGGTTTCACCAGGTTGGCCAGGCTGGTCTTGAACTCCTGACTTCAGGTGATCTGCCACCTAGGCCTCCCAAAGTGCTAGGATTACAG

GCGTGAGCCACTCTGCCCGGCCTCCTTCCTTCCTTCCTTTCTTTCTTTTTCTTCCTTCCTTCTTTCCTTTCTTTCTCTCTCTCTCTCTCT

CTGCCTCTTTCAGACAGAGTCAGGCACTGTTGCCTGGGCTGGAGTGCAATAGCATGATCTCAGCTCACTGCAACCTCCGCCTCCCAGGTT

CATGCGATTATCCTGCC

REN90965(F8Int24.1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 287 bp

Sequence 2: ref|AC_000066.1| 287 bp

Sequence 3: ref|AC_000155.1| 287 bp

ref|NC_000023.9| ATGAGGTTTCACCAGGTTGGCCAGGCTGGTCTTGAACTCCTGACTTCAGGTGATCTGCCA

ref|AC_000155.1| ATGAGGTTTCACCAGGTTGGCCAGGCTGGTCTTGAACTCCTGACTTCAGGTGATCTGCCA

ref|AC_000066.1| ATGAGGTTTCACCAGGTTGGCCAGGCTGGTCTTGAACTCCTGACTTCAGGTGATCTGCCA

************************************************************

ref|NC_000023.9| CCTAGGCCTCCCAAAGTGCTAGGATTACAGGCGTGAGCCACTCTGCCCGGCCTCCTTCCT

ref|AC_000155.1| CCTAGGCCTCCCAAAGTGCTAGGATTACAGGCGTGAGCCACTCTGCCCGGCCTCCTTCCT

ref|AC_000066.1| CCTAGGCCTCCCAAAGTGCTAGGATTACAGGCGTGAGCCACTCTGCCCGGCCTCCTTCCT

************************************************************

ref|NC_000023.9| TCCTTCCTTTCTTTCTTTTTCTTCCTTCCTTCTTTCCTTTCTTTCTCTCTCTCTCTCTCT

ref|AC_000155.1| TCCTTCCTTTCTTTCTTTTTCTTCCTTCCTTCTTTCCTTTCTTTCTCTCTCTCTCTCTCT

ref|AC_000066.1| TCCTTCCTTTCTTTCTTTTTCTTCCTTCCTTCTTTCCTTTCTTTCTCTCTCTCTCTCTCT

************************************************************

ref|NC_000023.9| CTGCCTCTTTCAGACAGAGTCAGGCACTGTTGCCTGGGCTGGAGTGCAATAGCATGATCT

ref|AC_000155.1| CTGCCTCTTTCAGACAGAGTCAGGCACTGTTGCCTGGGCTGGAGTGCAATAGCATGATCT

ref|AC_000066.1| CTGCCTCTTTCAGACAGAGTCAGGCACTGTTGCCTGGGCTGGAGTGCAATAGCATGATCT

************************************************************

ref|NC_000023.9| CAGCTCACTGCAACCTCCGCCTCCCAGGTTCATGCGATTATCCTGCC

ref|AC_000155.1| CAGCTCACTGCAACCTCCGCCTCCCAGGTTCATGCGATTATCCTGCC

ref|AC_000066.1| CAGCTCACTGCAACCTCCGCCTCCCAGGTTCATGCGATTATCCTGCC

***********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
F8Int22	NC_000023	153757206--153757243	19.0	100	76
	AC_000066	154262633--154262668	18.0	100	72
	AC_000155	142647511--142647548	19.0	100	76

New!	This microsatellite marker has been recently validated experimentally as informative^[1,2].
New!	Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[3]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect).
New!	This locus has recently been tried-and-tested as informative using the pair of primers here provided^[4].
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[5]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.52 and 0.55, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.
^[2]	de Sá Machado Araújo G, Louvain de Souza T, Medina-Acosta E. Use of SNPSTR combined markers for gametic phase determination of X chromosomes in Hemophilia A female carriers. In 58o. Congresso Brasileiro de Genética. 2012. Foz do Iguaçu: Sociedade Brasileira de Genética.
^[3]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.
^[4]	Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.
^[5]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

F8Int22	Information
Amplicon	221-223 bp
Forward ampliprimer	AAGACCCTTAGCTGTTTCATAAGC
Reverse ampliprimer	TTCATACAGTGGGATCATTCATT

>ref|NC_000023.9|

AAGACCCTTAGCTGTTTCATAAGCAGTAAAGTTTGAAAACACTGGCAGAGGACACAGTCAGCTGTGAAAGAGTACTGGGAATGCACAGCC

TATCCTCCACCCTGGTCTCAGAATGATCTTCTAAGAATGTAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAGAGAGAGA

GAGTCTATAATGTGGGCATTAATGAATGATCCCACTGTATGAA

>ref|AC_000066.1|

AAGACCCTTAGCTGTTTCATAAGCAGTAAAGTTTGAAAACACTGGCAGAGGACACAGTCAGCTGTGAAAGAGTACTGGGAATGCACAGCC

TATCCTCCACCCTGGTCTCAGAATGATCTTCTAAGAATGTAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAGAGAGAGAGA

GTCTATAATGTGGGCATTAATGAATGATCCCACTGTATGAA

>ref|AC_000155.1|

AAGACCCTTAGCTGTTTCATAAGCAGTAAAGTTTGAAAACACTGGCAGAGGACACAGTCAGCTGTGAAAGAGTACTGGGAATGCACAGCC

TATCCTCCACCCTGGTCTCAGAATGATCTTCTAAGAATGTAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAGAGAGAGA

GAGTCTATAATGTGGGCATTAATGAATGATCCCACTGTATGAA

F8Int22

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 223 bp

Sequence 2: ref|AC_000066.1| 221 bp

Sequence 3: ref|AC_000155.1| 223 bp

ref|NC_000023.9| AAGACCCTTAGCTGTTTCATAAGCAGTAAAGTTTGAAAACACTGGCAGAGGACACAGTCA

ref|AC_000155.1| AAGACCCTTAGCTGTTTCATAAGCAGTAAAGTTTGAAAACACTGGCAGAGGACACAGTCA

ref|AC_000066.1| AAGACCCTTAGCTGTTTCATAAGCAGTAAAGTTTGAAAACACTGGCAGAGGACACAGTCA

************************************************************

ref|NC_000023.9| GCTGTGAAAGAGTACTGGGAATGCACAGCCTATCCTCCACCCTGGTCTCAGAATGATCTT

ref|AC_000155.1| GCTGTGAAAGAGTACTGGGAATGCACAGCCTATCCTCCACCCTGGTCTCAGAATGATCTT

ref|AC_000066.1| GCTGTGAAAGAGTACTGGGAATGCACAGCCTATCCTCCACCCTGGTCTCAGAATGATCTT

************************************************************

ref|NC_000023.9| CTAAGAATGTAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAGAGAGAGA

ref|AC_000155.1| CTAAGAATGTAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAGAGAGAGA

ref|AC_000066.1| CTAAGAATGTAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGT--AGAGAGAGAGA

*********************************************** ***********

ref|NC_000023.9| GAGTCTATAATGTGGGCATTAATGAATGATCCCACTGTATGAA

ref|AC_000155.1| GAGTCTATAATGTGGGCATTAATGAATGATCCCACTGTATGAA

ref|AC_000066.1| GAGTCTATAATGTGGGCATTAATGAATGATCCCACTGTATGAA

*******************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HSC271_(M4)/F8Int21	NC_000023	153777976--153778027	26.0	92	86
	AC_000066	154283586--154283637	26.0	92	86
	AC_000155	142669525--142669576	26.0	92	86

New!	This microsatellite marker has been recently validated experimentally as informative ^[1,2].
New!	Multiallelic interlocus non-random association analysis revealed that **GAB3Int1 at gametic (linkage) EQUILIBRIUM with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1*.1. The observed breakdown of gametic disequilibrium attests historical recombination between GAB3*Int1 and the F8 gene.
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[3]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.48 and 0.51, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Alves Da Silva AF, Rossetti LC, De Brasi CD, Medina-Acosta E. Informativeness of a novel multiallelic marker-set comprising an F8 intron 21 and three tightly linked loci for haemophilia A carriership analysis. Haemophilia 2011, 17:257-66.
^[2]	de Sá Machado Araújo G, Louvain de Souza T, Medina-Acosta E. Use of SNPSTR combined markers for gametic phase determination of X chromosomes in Hemophilia A female carriers. In 58o. Congresso Brasileiro de Genética. 2012. Foz do Iguaçu: Sociedade Brasileira de Genética.
^[3]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

HSC271_(M4)/F8Int21	Information
Amplicon	164 bp
Forward ampliprimer	AGCCTGCTTCTTGACAAACC
Reverse ampliprimer	AGGTGTGGGCAAAGCATTAG

>ref|NC_000023.9|

AGCCTGCTTCTTGACAAACCAAACCACTACCAAATCATTGAGCCCCCATTTCGTACACACACACACACACACACACACACACACATACAC

ACAAACACACACACACCTTTTGTGGACTCACATAAGCTAATGGACCCACATAAGCTAATGCTTTGCCCACACCT

>ref|AC_000066.1|

AGCCTGCTTCTTGACAAACCAAACCACTACCAAATCATTGAGCCCCCATTTCGTACACACACACACACACACACACACACACACATACAC

ACAAACACACACACACCTTTTGTGGACTCACATAAGCTAATGGACCCACATAAGCTAATGCTTTGCCCACACCT

>ref|AC_000155.1|

AGCCTGCTTCTTGACAAACCAAACCACTACCAAATCATTGAGCCCCCATTTCGTACACACACACACACACACACACACACACACATACAC

ACAAACACACACACACCTTTTGTGGACTCACATAAGCTAATGGACCCACATAAGCTAATGCTTTGCCCACACCT

HSC271_(M4)/F8Int21

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 164 bp

Sequence 2: ref|AC_000066.1| 164 bp

Sequence 3: ref|AC_000155.1| 164 bp

ref|NC_000023.9| AGCCTGCTTCTTGACAAACCAAACCACTACCAAATCATTGAGCCCCCATTTCGTACACAC

ref|AC_000155.1| AGCCTGCTTCTTGACAAACCAAACCACTACCAAATCATTGAGCCCCCATTTCGTACACAC

ref|AC_000066.1| AGCCTGCTTCTTGACAAACCAAACCACTACCAAATCATTGAGCCCCCATTTCGTACACAC

************************************************************

ref|NC_000023.9| ACACACACACACACACACACACACATACACACAAACACACACACACCTTTTGTGGACTCA

ref|AC_000155.1| ACACACACACACACACACACACACATACACACAAACACACACACACCTTTTGTGGACTCA

ref|AC_000066.1| ACACACACACACACACACACACACATACACACAAACACACACACACCTTTTGTGGACTCA

************************************************************

ref|NC_000023.9| CATAAGCTAATGGACCCACATAAGCTAATGCTTTGCCCACACCT

ref|AC_000155.1| CATAAGCTAATGGACCCACATAAGCTAATGCTTTGCCCACACCT

ref|AC_000066.1| CATAAGCTAATGGACCCACATAAGCTAATGCTTTGCCCACACCT

********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN91100(F8Int13.2)	NC_000023	153817451--153817493	21.5	100	86
	AC_000066	154322995--154323035	20.5	100	82
	AC_000155	142709064--142709104	20.5	100	82

New!	This microsatellite marker has been recently validated experimentally as informative^[1].
New!	Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[2]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect).
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[3]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.63 and 0.57, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.
^[2]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.
^[3]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

REN91100(F8Int13.2)	Information
Amplicon	148-150 bp
Forward ampliprimer	GCCTAGAGAATGCCAAAGTAACA
Reverse ampliprimer	TTGTTGTAATTCCCATTTGCAT

>ref|NC_000023.9|

GCCTAGAGAATGCCAAAGTAACACACACACACACACACACACACACACACACACACACACACAAAAATCAAAGTAAAGAGCTTGAAGCCA

TCCAGATTTTTTTAAAAAGATACATTATGTACAGTTGAATGCAAATGGGAATTACAACAA

>ref|AC_000066.1|

GCCTAGAGAATGCCAAAGTAACACACACACACACACACACACACACACACACACACACACAAAAATCAAAGTAAAGAGCTTGAAGCCATC

CAGATTTTTTTAAAAAGATACATTATGTACAGTTGAATGCAAATGGGAATTACAACAA

>ref|AC_000155.1|

GCCTAGAGAATGCCAAAGTAACACACACACACACACACACACACACACACACACACACACAAAAATCAAAGTAAAGAGCTTGAAGCCATC

CAGATTTTTTTAAAAAGATACATTATGTACAGTTGAATGCAAATGGGAATTACAACAA

REN91100(F8Int13.2)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 150 bp

Sequence 2: ref|AC_000066.1| 148 bp

Sequence 3: ref|AC_000155.1| 148 bp

ref|AC_000066.1| GCCTAGAGAATGCCAAAGTAACACACACACACACACACACACACACACACACACACACAC

ref|AC_000155.1| GCCTAGAGAATGCCAAAGTAACACACACACACACACACACACACACACACACACACACAC

ref|NC_000023.9| GCCTAGAGAATGCCAAAGTAACACACACACACACACACACACACACACACACACACACAC

************************************************************

ref|AC_000066.1| A--AAAATCAAAGTAAAGAGCTTGAAGCCATCCAGATTTTTTTAAAAAGATACATTATGT

ref|AC_000155.1| A--AAAATCAAAGTAAAGAGCTTGAAGCCATCCAGATTTTTTTAAAAAGATACATTATGT

ref|NC_000023.9| ACAAAAATCAAAGTAAAGAGCTTGAAGCCATCCAGATTTTTTTAAAAAGATACATTATGT

* *********************************************************

ref|AC_000066.1| ACAGTTGAATGCAAATGGGAATTACAACAA

ref|AC_000155.1| ACAGTTGAATGCAAATGGGAATTACAACAA

ref|NC_000023.9| ACAGTTGAATGCAAATGGGAATTACAACAA

******************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN91114(F8Int13.1)	NC_000023	153820354--153820370	8.5	100	34
	AC_000066	154325897--154325915	9.5	100	38
	AC_000155	142711931--142711947	8.5	100	34

REN91114(F8Int13.1)	Information
Amplicon	264-266 bp
Forward ampliprimer	GCCAACACACACTGAAGGAA
Reverse ampliprimer	CTCCCAATGCAAAGACCTTC

>ref|NC_000023.9|

GCCAACACACACTGAAGGAAGTTCCAAAGAAGTTTCTTCAGACAAAAAGAAAATGATCATAGATGGAAGCAAGAAATGCAGAAAAGAATG

AAAGAAATGAAAGGTGTAAATATATATATATATATAAATGATATTGATTATATAAAACAATAATAATGTCTTATGGGGTTTAAAATATAT

GTAGAATTCAAATAAATGACAAGAGCACAAAAGGTAGGAGTGAGAAAATGAAGTCACGTTGTTTGAAGGTCTTTGCATTGGGAG

>ref|AC_000066.1|

GCCAACACACACTGAAGGAAGTTCCAAAGAAGTTTCTTCAGACAAAAAGAAAATGATCATAGATGGAAGCAAGAAATGCAGAAAAGAATG

AAAGAAATGAAAGGTGTAAATATATATATATATATATAAATGATATTGATTATATAAAACAATAATAATGTCTTATGGGGTTTAAAATAT

ATGTAGAATTCAAATAAATGACAAGAGCACAAAAGGTAGGAGTGAGAAAATGAAGTCACGTTGTTTGAAGGTCTTTGCATTGGGAG

>ref|AC_000155.1|

GCCAACACACACTGAAGGAAGTTCCAAAGAAGTTTCTTCAGACAAAAAGAAAATGATCATAGATGGAAGCAAGAAATGCAGAAAAGAATG

AAAGAAATGAAAGGTGTAAATATATATATATATATAAATGATATTGATTATATAAAACAATAATAATGTCTTATGGGGTTTAAAATATAT

GTAGAATTCAAATAAATGACAAGAGCACAAAAGGTAGGAGTGAGAAAATGAAGTCACGTTGTTTGAAGGTCTTTGCATTGGGAG

REN91114(F8Int13.1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 264 bp

Sequence 2: ref|AC_000066.1| 266 bp

Sequence 3: ref|AC_000155.1| 264 bp

ref|NC_000023.9| GCCAACACACACTGAAGGAAGTTCCAAAGAAGTTTCTTCAGACAAAAAGAAAATGATCAT

ref|AC_000155.1| GCCAACACACACTGAAGGAAGTTCCAAAGAAGTTTCTTCAGACAAAAAGAAAATGATCAT

ref|AC_000066.1| GCCAACACACACTGAAGGAAGTTCCAAAGAAGTTTCTTCAGACAAAAAGAAAATGATCAT

************************************************************

ref|NC_000023.9| AGATGGAAGCAAGAAATGCAGAAAAGAATGAAAGAAATGAAAGGTGTAAATATATATATA

ref|AC_000155.1| AGATGGAAGCAAGAAATGCAGAAAAGAATGAAAGAAATGAAAGGTGTAAATATATATATA

ref|AC_000066.1| AGATGGAAGCAAGAAATGCAGAAAAGAATGAAAGAAATGAAAGGTGTAAATATATATATA

************************************************************

ref|NC_000023.9| TATATA--AATGATATTGATTATATAAAACAATAATAATGTCTTATGGGGTTTAAAATAT

ref|AC_000155.1| TATATA--AATGATATTGATTATATAAAACAATAATAATGTCTTATGGGGTTTAAAATAT

ref|AC_000066.1| TATATATAAATGATATTGATTATATAAAACAATAATAATGTCTTATGGGGTTTAAAATAT

****** ****************************************************

ref|NC_000023.9| ATGTAGAATTCAAATAAATGACAAGAGCACAAAAGGTAGGAGTGAGAAAATGAAGTCACG

ref|AC_000155.1| ATGTAGAATTCAAATAAATGACAAGAGCACAAAAGGTAGGAGTGAGAAAATGAAGTCACG

ref|AC_000066.1| ATGTAGAATTCAAATAAATGACAAGAGCACAAAAGGTAGGAGTGAGAAAATGAAGTCACG

************************************************************

ref|NC_000023.9| TTGTTTGAAGGTCTTTGCATTGGGAG

ref|AC_000155.1| TTGTTTGAAGGTCTTTGCATTGGGAG

ref|AC_000066.1| TTGTTTGAAGGTCTTTGCATTGGGAG

**************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604366/REN91179/REN91180(F8Int10)	NC_000023	153842062--153842082	10.5	100	42
	AC_000066	154347607--154347627	10.5	100	42
	AC_000155	142733619--142733639	10.5	100	42

stSG604366/REN91179/REN91180(F8Int10)	Information
Amplicon	201 bp
Forward ampliprimer	TGAATATGCGAAGAACCCTGA
Reverse ampliprimer	CGGTTTTCAGCTACATTCCAC

>ref|NC_000023.9|

TGAATATGCGAAGAACCCTGAAAATATCAAACTAGATACCAAAAGTGAGAGTGAAACCCTGTTGAGAGAGAGAGAGAGAGAGAGCAAGGA

AGAGCAATAACATGGAAGCTGCAGCAAAGAATCCTGAAAATATCAAACTAGTTACCCAAAGTGTGGAATACCACACTAGATACCAAAAGT

GTGGAATGTAGCTGAAAACCG

>ref|AC_000066.1|

TGAATATGCGAAGAACCCTGAAAATATCAAACTAGATACCAAAAGTGAGAGTGAAACCCTGTTGAGAGAGAGAGAGAGAGAGAGCAAGGA

AGAGCAATAACATGGAAGCTGCAGCAAAGAATCCTGAAAATATCAAACTAGTTACCCAAAGTGTGGAATACCACACTAGATACCAAAAGT

GTGGAATGTAGCTGAAAACCG

>ref|AC_000155.1|

TGAATATGCGAAGAACCCTGAAAATATCAAACTAGATACCAAAAGTGAGAGTGAAACCCTGTTGAGAGAGAGAGAGAGAGAGAGCAAGGA

AGAGCAATAACATGGAAGCTGCAGCAAAGAATCCTGAAAATATCAAACTAGTTACCCAAAGTGTGGAATACCACACTAGATACCAAAAGT

GTGGAATGTAGCTGAAAACCG

stSG604366/REN91179/REN91180(F8Int10)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 201 bp

Sequence 2: ref|AC_000066.1| 201 bp

Sequence 3: ref|AC_000155.1| 201 bp

ref|NC_000023.9| TGAATATGCGAAGAACCCTGAAAATATCAAACTAGATACCAAAAGTGAGAGTGAAACCCT

ref|AC_000155.1| TGAATATGCGAAGAACCCTGAAAATATCAAACTAGATACCAAAAGTGAGAGTGAAACCCT

ref|AC_000066.1| TGAATATGCGAAGAACCCTGAAAATATCAAACTAGATACCAAAAGTGAGAGTGAAACCCT

************************************************************

ref|NC_000023.9| GTTGAGAGAGAGAGAGAGAGAGAGCAAGGAAGAGCAATAACATGGAAGCTGCAGCAAAGA

ref|AC_000155.1| GTTGAGAGAGAGAGAGAGAGAGAGCAAGGAAGAGCAATAACATGGAAGCTGCAGCAAAGA

ref|AC_000066.1| GTTGAGAGAGAGAGAGAGAGAGAGCAAGGAAGAGCAATAACATGGAAGCTGCAGCAAAGA

************************************************************

ref|NC_000023.9| ATCCTGAAAATATCAAACTAGTTACCCAAAGTGTGGAATACCACACTAGATACCAAAAGT

ref|AC_000155.1| ATCCTGAAAATATCAAACTAGTTACCCAAAGTGTGGAATACCACACTAGATACCAAAAGT

ref|AC_000066.1| ATCCTGAAAATATCAAACTAGTTACCCAAAGTGTGGAATACCACACTAGATACCAAAAGT

************************************************************

ref|NC_000023.9| GTGGAATGTAGCTGAAAACCG

ref|AC_000155.1| GTGGAATGTAGCTGAAAACCG

ref|AC_000066.1| GTGGAATGTAGCTGAAAACCG

*********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604368(F8Int9.2)	NC_000023	153844339--153844390	25.5	92	86
	AC_000066	154349884--154349935	25.5	92	86
	AC_000155	142735826--142735877	25.5	92	86

New!	This microsatellite marker has been recently validated experimentally as informative ^[1].
New!	Multiallelic interlocus non-random association analysis revealed that **GAB3Int1 is at gametic (linkage) EQUILIBRIUM with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1*.1. The observed breakdown of gametic disequilibrium attests historical recombination between GAB3*Int1 and the F8 gene.
^[1]	Machado FB, Alves Da Silva AF, Rossetti LC, De Brasi CD, Medina-Acosta E. Informativeness of a novel multiallelic marker-set comprising an F8 intron 21 and three tightly linked loci for haemophilia A carriership analysis. Haemophilia 2011, 17:257-66.

stSG604368(F8Int9.2)	Information
Amplicon	165-166 bp
Forward ampliprimer	CTGGGTGACAGAATGAGACC
Reverse ampliprimer	TCCTCATATCACACACCTCAGA

>ref|NC_000023.9|

CTGGGTGACAGAATGAGACCCTATCTCAAGAGAGAGAGAGAGAGACAGAGACGAGAGAGAGAGAGAGAGAGAGAGAGAGAATGGAAGAGC

AATAACATGGAAGCTGCAGCGAAGAACCCTGAAAATATCAATCCCACCTCCAGATCTGAGGTGTGTGATATGAGGA

>ref|AC_000066.1|

CTGGGTGACAGAATGAGACCCTATCTCAAGAGAGAGAGAGAGAGACAGAGACGAGAGAGAGAGAGAGAGAGAGAGAGAGAATGGAAGAGC

AATAACATGGAAGCTGCAGCGAAGAACCCTGAAAATATCAATCCCACCTCCAGATCTGAGGTGTGTGATATGAGGA

>ref|AC_000155.1|

CTGGGTGACAGAATGAGACCCTATCTCAGAGAGAGAGAGAGAGACAGAGACGAGAGAGAGAGAGAGAGAGAGAGAGAGAATGGAAGAGCA

ATAACATGGAAGCTGCAGCGAAGAACCCTGAAAATATCAATCCCACCTCCAGATCTGAGGTGTGTGATATGAGGA

stSG604368(F8Int9.2)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 166 bp

Sequence 2: ref|AC_000066.1| 166 bp

Sequence 3: ref|AC_000155.1| 165 bp

ref|NC_000023.9| CTGGGTGACAGAATGAGACCCTATCTCAAGAGAGAGAGAGAGAGACAGAGACGAGAGAGA

ref|AC_000066.1| CTGGGTGACAGAATGAGACCCTATCTCAAGAGAGAGAGAGAGAGACAGAGACGAGAGAGA

ref|AC_000155.1| CTGGGTGACAGAATGAGACCCTATCTCA-GAGAGAGAGAGAGAGACAGAGACGAGAGAGA

**************************** *******************************

ref|NC_000023.9| GAGAGAGAGAGAGAGAGAGAATGGAAGAGCAATAACATGGAAGCTGCAGCGAAGAACCCT

ref|AC_000066.1| GAGAGAGAGAGAGAGAGAGAATGGAAGAGCAATAACATGGAAGCTGCAGCGAAGAACCCT

ref|AC_000155.1| GAGAGAGAGAGAGAGAGAGAATGGAAGAGCAATAACATGGAAGCTGCAGCGAAGAACCCT

************************************************************

ref|NC_000023.9| GAAAATATCAATCCCACCTCCAGATCTGAGGTGTGTGATATGAGGA

ref|AC_000066.1| GAAAATATCAATCCCACCTCCAGATCTGAGGTGTGTGATATGAGGA

ref|AC_000155.1| GAAAATATCAATCCCACCTCCAGATCTGAGGTGTGTGATATGAGGA

**********************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604369/REN91192(F8Int9.1)	NC_000023	153845551--153845578	14.0	92	47
	AC_000066	154351096--154351123	14.0	92	47
	AC_000155	142737038--142737065	14.0	92	47

New!

This locus has recently been tried-and-tested as informative using the pair of primers here provided^[1].

^[1]

Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.

stSG604369/REN91192(F8Int9.1)	Information
Amplicon	233 bp
Forward ampliprimer	AGATTCGAGCGATTCTCCTG
Reverse ampliprimer	CAGTCATTGCTGTGGGTTTG

>ref|NC_000023.9|

AGATTCGAGCGATTCTCCTGCCTCAGCCTCCAGAGTAGCTGGGACTACAGGCATGTGCCACCATGCCTGGCTAATTCAAAATATATTGTT

ACCTTTTAAAAATCAAGGTACAGACCAATTTACACTGCTCTTGTTTTTGTGTGAAATGCACACACACACACACATACACACACACATTTG

TACATGTATGCACTACTTCTGGAGGGGAGTATACAAACCCACAGCAATGACTG

>ref|AC_000066.1|

AGATTCGAGCGATTCTCCTGCCTCAGCCTCCAGAGTAGCTGGGACTACAGGCATGTGCCACCATGCCTGGCTAATTCAAAATATATTGTT

ACCTTTTAAAAATCAAGGTACAGACCAATTTACACTGCTCTTGTTTTTGTGTGAAATGCACACACACACACACATACACACACACATTTG

TACATGTATGCACTACTTCTGGAGGGGAGTATACAAACCCACAGCAATGACTG

>ref|AC_000155.1|

AGATTCGAGCGATTCTCCTGCCTCAGCCTCCAGAGTAGCTGGGACTACAGGCATGTGCCACCATGCCTGGCTAATTCAAAATATATTGTT

ACCTTTTAAAAATCAAGGTACAGACCAATTTACACTGCTCTTGTTTTTGTGTGAAATGCACACACACACACACATACACACACACATTTG

TACATGTATGCACTACTTCTGGAGGGGAGTATACAAACCCACAGCAATGACTG

stSG604369/REN91192(F8Int9.1)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 233 bp

Sequence 2: ref|AC_000066.1| 233 bp

Sequence 3: ref|AC_000155.1| 233 bp

ref|NC_000023.9| AGATTCGAGCGATTCTCCTGCCTCAGCCTCCAGAGTAGCTGGGACTACAGGCATGTGCCA

ref|AC_000155.1| AGATTCGAGCGATTCTCCTGCCTCAGCCTCCAGAGTAGCTGGGACTACAGGCATGTGCCA

ref|AC_000066.1| AGATTCGAGCGATTCTCCTGCCTCAGCCTCCAGAGTAGCTGGGACTACAGGCATGTGCCA

************************************************************

ref|NC_000023.9| CCATGCCTGGCTAATTCAAAATATATTGTTACCTTTTAAAAATCAAGGTACAGACCAATT

ref|AC_000155.1| CCATGCCTGGCTAATTCAAAATATATTGTTACCTTTTAAAAATCAAGGTACAGACCAATT

ref|AC_000066.1| CCATGCCTGGCTAATTCAAAATATATTGTTACCTTTTAAAAATCAAGGTACAGACCAATT

************************************************************

ref|NC_000023.9| TACACTGCTCTTGTTTTTGTGTGAAATGCACACACACACACACATACACACACACATTTG

ref|AC_000155.1| TACACTGCTCTTGTTTTTGTGTGAAATGCACACACACACACACATACACACACACATTTG

ref|AC_000066.1| TACACTGCTCTTGTTTTTGTGTGAAATGCACACACACACACACATACACACACACATTTG

************************************************************

ref|NC_000023.9| TACATGTATGCACTACTTCTGGAGGGGAGTATACAAACCCACAGCAATGACTG

ref|AC_000155.1| TACATGTATGCACTACTTCTGGAGGGGAGTATACAAACCCACAGCAATGACTG

ref|AC_000066.1| TACATGTATGCACTACTTCTGGAGGGGAGTATACAAACCCACAGCAATGACTG

*****************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
REN91235(F8Int6)	NC_000023	153855435--153855461	13.5	100	54
	AC_000066	154360978--154361004	13.5	100	54
	AC_000155	142747376--142747402	13.5	100	54

New!

This locus has recently been tried-and-tested as informative using the pair of primers here provided^[1].

^[1]

Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.

REN91235(F8Int6)	Information
Amplicon	325 bp
Forward ampliprimer	TTCTCCTGCTTCAGCCTCTC
Reverse ampliprimer	AGCATATCCACCCTCACCAC

>ref|NC_000023.9|

TTCTCCTGCTTCAGCCTCTCGAGTAGCTGGGACTACAGGAGCGCACCACCATGTCTGGCTAGCATGTGTGTGTGTGTGTGTGTGTGTGTG

TATTTTTAGTAGAGATGGGGTTTCACCATATTGCCAGGCTGGTCTCGAACTCTGGACATCAAGTGATCTGCCCGCCTTGGCCTCCCAAAG

TTCTGGGATTACAGGCATAAGCCATTGAGCCTGGCCAAACAGGGACAATTTGACTTCCTTCTTTCCAATTTGGATGTCTTGCCTAATTGC

TCTGGCTAGGACTTCCAGTACTGTGTTGAATAGAAGTGGTGAGGGTGGATATGCT

>ref|AC_000066.1|

TTCTCCTGCTTCAGCCTCTCGAGTAGCTGGGACTACAGGAGCGCACCACCATGTCTGGCTAGCATGTGTGTGTGTGTGTGTGTGTGTGTG

TATTTTTAGTAGAGATGGGGTTTCACCATATTGCCAGGCTGGTCTCGAACTCTGGACATCAAGTGATCTGCCCGCCTTGGCCTCCCAAAG

TTCTGGGATTACAGGCATAAGCCATTGAGCCTGGCCAAACAGGGACAATTTGACTTCCTTCTTTCCAATTTGGATGTCTTGCCTAATTGC

TCTGGCTAGGACTTCCAGTACTGTGTTGAATAGAAGTGGTGAGGGTGGATATGCT

>ref|AC_000155.1|

TTCTCCTGCTTCAGCCTCTCGAGTAGCTGGGACTACAGGAGCGCACCACCATGTCTGGCTAGCATGTGTGTGTGTGTGTGTGTGTGTGTG

TATTTTTAGTAGAGATGGGGTTTCACCATATTGCCAGGCTGGTCTCGAACTCTGGACATCAAGTGATCTGCCCGCCTTGGCCTCCCAAAG

TTCTGGGATTACAGGCATAAGCCATTGAGCCTGGCCAAACAGGGACAATTTGACTTCCTTCTTTCCAATTTGGATGTCTTGCCTAATTGC

TCTGGCTAGGACTTCCAGTACTGTGTTGAATAGAAGTGGTGAGGGTGGATATGCT

REN91235(F8Int6)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 325 bp

Sequence 2: ref|AC_000066.1| 325 bp

Sequence 3: ref|AC_000155.1| 325 bp

ref|AC_000066.1| TTCTCCTGCTTCAGCCTCTCGAGTAGCTGGGACTACAGGAGCGCACCACCATGTCTGGCT

ref|NC_000023.9| TTCTCCTGCTTCAGCCTCTCGAGTAGCTGGGACTACAGGAGCGCACCACCATGTCTGGCT

ref|AC_000155.1| TTCTCCTGCTTCAGCCTCTCGAGTAGCTGGGACTACAGGAGCGCACCACCATGTCTGGCT

************************************************************

ref|AC_000066.1| AGCATGTGTGTGTGTGTGTGTGTGTGTGTGTATTTTTAGTAGAGATGGGGTTTCACCATA

ref|NC_000023.9| AGCATGTGTGTGTGTGTGTGTGTGTGTGTGTATTTTTAGTAGAGATGGGGTTTCACCATA

ref|AC_000155.1| AGCATGTGTGTGTGTGTGTGTGTGTGTGTGTATTTTTAGTAGAGATGGGGTTTCACCATA

************************************************************

ref|AC_000066.1| TTGCCAGGCTGGTCTCGAACTCTGGACATCAAGTGATCTGCCCGCCTTGGCCTCCCAAAG

ref|NC_000023.9| TTGCCAGGCTGGTCTCGAACTCTGGACATCAAGTGATCTGCCCGCCTTGGCCTCCCAAAG

ref|AC_000155.1| TTGCCAGGCTGGTCTCGAACTCTGGACATCAAGTGATCTGCCCGCCTTGGCCTCCCAAAG

************************************************************

ref|AC_000066.1| TTCTGGGATTACAGGCATAAGCCATTGAGCCTGGCCAAACAGGGACAATTTGACTTCCTT

ref|NC_000023.9| TTCTGGGATTACAGGCATAAGCCATTGAGCCTGGCCAAACAGGGACAATTTGACTTCCTT

ref|AC_000155.1| TTCTGGGATTACAGGCATAAGCCATTGAGCCTGGCCAAACAGGGACAATTTGACTTCCTT

************************************************************

ref|AC_000066.1| CTTTCCAATTTGGATGTCTTGCCTAATTGCTCTGGCTAGGACTTCCAGTACTGTGTTGAA

ref|NC_000023.9| CTTTCCAATTTGGATGTCTTGCCTAATTGCTCTGGCTAGGACTTCCAGTACTGTGTTGAA

ref|AC_000155.1| CTTTCCAATTTGGATGTCTTGCCTAATTGCTCTGGCTAGGACTTCCAGTACTGTGTTGAA

************************************************************

ref|AC_000066.1| TAGAAGTGGTGAGGGTGGATATGCT

ref|NC_000023.9| TAGAAGTGGTGAGGGTGGATATGCT

ref|AC_000155.1| TAGAAGTGGTGAGGGTGGATATGCT

*************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604384/REN91318(F8Int3)	NC_000023	153874731--153874753	7.7	100	46
	AC_000066	154380272--154380294	7.7	100	46
	AC_000155	142766684--142766706	7.7	100	46

stSG604384/REN91318(F8Int3)	Information
Amplicon	221 bp
Forward ampliprimer	AGCAGGAAGAAATAAAATTGTCC
Reverse ampliprimer	CTGAGGCAGGAAAATCACTTG

>ref|NC_000023.9|

AGCAGGAAGAAATAAAATTGTCCTTTCTATATCCACTGTACACTCAAAGAAACATGTCAAGTTTATTGTCACCAATAGGTTCTACATAAG

AAACGGACTTTCTGCATCTTTGTTGTTGTTGTTGTTGTTGTTTTGAGACGGAGTCTCACTCTGTTGCCCAGGCTGGATCTCGGCTCACAG

CAACGTCCGCCTCCCAGGTTCAAGTGATTTTCCTGCCTCAG

>ref|AC_000066.1|

AGCAGGAAGAAATAAAATTGTCCTTTCTATATCCACTGTACACTCAAAGAAACATGTCAAGTTTATTGTCACCAATAGGTTCTACATAAG

AAACGGACTTTCTGCATCTTTGTTGTTGTTGTTGTTGTTGTTTTGAGACGGAGTCTCACTCTGTTGCCCAGGCTGGATCTCGGCTCACAG

CAACGTCCGCCTCCCAGGTTCAAGTGATTTTCCTGCCTCAG

>ref|AC_000155.1|

AGCAGGAAGAAATAAAATTGTCCTTTCTATATCCACTGTACACTCAAAGAAACATGTCAAGTTTATTGTCACCAATAGGTTCTACATAAG

AAACGGACTTTCTGCATCTTTGTTGTTGTTGTTGTTGTTGTTTTGAGACGGAGTCTCACTCTGTTGCCCAGGCTGGATCTCGGCTCACAG

CAACGTCCGCCTCCCAGGTTCAAGTGATTTTCCTGCCTCAG

stSG604384/REN91318(F8Int3)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 221 bp

Sequence 2: ref|AC_000066.1| 221 bp

Sequence 3: ref|AC_000155.1| 221 bp

ref|NC_000023.9| AGCAGGAAGAAATAAAATTGTCCTTTCTATATCCACTGTACACTCAAAGAAACATGTCAA

ref|AC_000155.1| AGCAGGAAGAAATAAAATTGTCCTTTCTATATCCACTGTACACTCAAAGAAACATGTCAA

ref|AC_000066.1| AGCAGGAAGAAATAAAATTGTCCTTTCTATATCCACTGTACACTCAAAGAAACATGTCAA

************************************************************

ref|NC_000023.9| GTTTATTGTCACCAATAGGTTCTACATAAGAAACGGACTTTCTGCATCTTTGTTGTTGTT

ref|AC_000155.1| GTTTATTGTCACCAATAGGTTCTACATAAGAAACGGACTTTCTGCATCTTTGTTGTTGTT

ref|AC_000066.1| GTTTATTGTCACCAATAGGTTCTACATAAGAAACGGACTTTCTGCATCTTTGTTGTTGTT

************************************************************

ref|NC_000023.9| GTTGTTGTTGTTTTGAGACGGAGTCTCACTCTGTTGCCCAGGCTGGATCTCGGCTCACAG

ref|AC_000155.1| GTTGTTGTTGTTTTGAGACGGAGTCTCACTCTGTTGCCCAGGCTGGATCTCGGCTCACAG

ref|AC_000066.1| GTTGTTGTTGTTTTGAGACGGAGTCTCACTCTGTTGCCCAGGCTGGATCTCGGCTCACAG

************************************************************

ref|NC_000023.9| CAACGTCCGCCTCCCAGGTTCAAGTGATTTTCCTGCCTCAG

ref|AC_000155.1| CAACGTCCGCCTCCCAGGTTCAAGTGATTTTCCTGCCTCAG

ref|AC_000066.1| CAACGTCCGCCTCCCAGGTTCAAGTGATTTTCCTGCCTCAG

*****************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
F8Int1	NC_000023	153884082--153884115	17.0	100	68
F8Int1	AC_000066	154389623--154389654	16.0	100	64

New!	This locus has recently been tried-and-tested as informative using the pair of ampliprimers here provided^[1].
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[2]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.45 and 0.43, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.
^[2]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

F8Int1	Information
Amplicon	218-220 bp
Forward ampliprimer	CTGCCCTTGGACATAAGCAT
Reverse ampliprimer	CCATATGATCCAGCAACTCG

>ref|NC_000023.9|

CTGCCCTTGGACATAAGCATTCTTGGTTCTGCCTTCTGACTCAGACCCTGTACTTTTACCATTGGCATATATTTACACACACACACACAC

ACACACACACACACACACGGATCTGTTTCATTGGAGAACCCTAATACAATTGATGTATAAATATCTCTTTAAGACCCCATTTCAGTTCTT

TTGCATATATAGCCAGAAGTCGAGTTGCTGGATCATATGG

>ref|AC_000066.1|

CTGCCCTTGGACATAAGCATTCTTGGTTCTGCCTTCTGACTCAGACCCTGTACTTTTACCATTGGCATATATTTACACACACACACACAC

ACACACACACACACACGGATCTGTTTCATTGGAGAACCCTAATACAATTGATGTATAAATATCTCTTTAAGACCCCATTTCAGTTCTTTT

GCATATATAGCCAGAAGTCGAGTTGCTGGATCATATGG

F8Int1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 220 bp

Sequence 2: ref|AC_000066.1| 218 bp

ref|NC_000023.9| CTGCCCTTGGACATAAGCATTCTTGGTTCTGCCTTCTGACTCAGACCCTGTACTTTTACC

ref|AC_000066.1| CTGCCCTTGGACATAAGCATTCTTGGTTCTGCCTTCTGACTCAGACCCTGTACTTTTACC

************************************************************

ref|NC_000023.9| ATTGGCATATATTTACACACACACACACACACACACACACACACACACGGATCTGTTTCA

ref|AC_000066.1| ATTGGCATATATTTACACACACACACACACACACACACACACACAC--GGATCTGTTTCA

********************************************** ************

ref|NC_000023.9| TTGGAGAACCCTAATACAATTGATGTATAAATATCTCTTTAAGACCCCATTTCAGTTCTT

ref|AC_000066.1| TTGGAGAACCCTAATACAATTGATGTATAAATATCTCTTTAAGACCCCATTTCAGTTCTT

************************************************************

ref|NC_000023.9| TTGCATATATAGCCAGAAGTCGAGTTGCTGGATCATATGG

ref|AC_000066.1| TTGCATATATAGCCAGAAGTCGAGTTGCTGGATCATATGG

****************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154006.7	NC_000023	154006702--154006794	46.5	91	96
	AC_000066	154512270--154512356	43.5	90	84
	AC_000155	142898749--142898835	43.5	90	84

HEMA154006.7	Information
Amplicon	274-276 bp
Forward ampliprimer	CATGGAAAGGAGGAAGGGTA
Reverse ampliprimer	TGGACGGAGCTGAGACAGAT

>ref|NC_000023.9|

CATGGAAAGGAGGAAGGGTAAAATGAATCTCATGGTATTGGATTAAAATTGGAAAAGAGTTCATTAATATGAACTCTTGTTCAAAGGAAA

TATATATATATATATATATACACACACACACACACACACACACATATATACACACACACACACATACACACACATATATACACACACACA

CACATATATACACACACACACAGAAACCCTATATATATATATCTCAGAGCTGAGAAAGGTATGTGTATATGTACATATCTGTCTCAGCTC

CGTCCA

>ref|AC_000066.1|

CATGGAAAGGAGGAAGGGTAAAATGAATCTCATGGTATTGGATTAAAATTGGAAAAGAGTTCATTAATATGAACTCTTGTTCAAAGGAAA

TATATATATATATATATATACATATACACACACACACACACACATATATACACACACACACACATACACACACATATATACACACACACA

CACATATATACACACACACACAGAAACCCTATATATATATATCTCAGAGCTGAGAAAGGTATGTGTATATGTACATATCTGTCTCAGCTC

CGTCCA

>ref|AC_000155.1|

CATGGAAAGGAGGAAGGGTAAAATGAATCTCATGGTATTGGATTAAAATTGGAAAAGAGTTCATTAATATGAACTCTTGTTCAAAGGAAA

TATATATATATATATATATATATACACACACACACACACACATATATACACACACACACACATACACACACATATATACACACACACACA

CATATATACACACACACACAGAAACCCTATATATATATATCTCAGAGCTGAGAAAGGTATGTGTATATGTACATATCTGTCTCAGCTCCG

TCCA

HEMA154006.7

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 276 bp

Sequence 2: ref|AC_000066.1| 276 bp

Sequence 3: ref|AC_000155.1| 274 bp

ref|NC_000023.9| CATGGAAAGGAGGAAGGGTAAAATGAATCTCATGGTATTGGATTAAAATTGGAAAAGAGT

ref|AC_000066.1| CATGGAAAGGAGGAAGGGTAAAATGAATCTCATGGTATTGGATTAAAATTGGAAAAGAGT

ref|AC_000155.1| CATGGAAAGGAGGAAGGGTAAAATGAATCTCATGGTATTGGATTAAAATTGGAAAAGAGT

************************************************************

ref|NC_000023.9| TCATTAATATGAACTCTTGTTCAAAGGAAATATATATATATATATATATACACACACACA

ref|AC_000066.1| TCATTAATATGAACTCTTGTTCAAAGGAAATATATATATATATATATATACATATACACA

ref|AC_000155.1| TCATTAATATGAACTCTTGTTCAAAGGAAATATATATATATATATATAT--ATATACACA

************************************************* * * *****

ref|NC_000023.9| CACACACACACACATATATACACACACACACACATACACACACATATATACACACACACA

ref|AC_000066.1| CACACACACACACATATATACACACACACACACATACACACACATATATACACACACACA

ref|AC_000155.1| CACACACACACACATATATACACACACACACACATACACACACATATATACACACACACA

************************************************************

ref|NC_000023.9| CACATATATACACACACACACAGAAACCCTATATATATATATCTCAGAGCTGAGAAAGGT

ref|AC_000066.1| CACATATATACACACACACACAGAAACCCTATATATATATATCTCAGAGCTGAGAAAGGT

ref|AC_000155.1| CACATATATACACACACACACAGAAACCCTATATATATATATCTCAGAGCTGAGAAAGGT

************************************************************

ref|NC_000023.9| ATGTGTATATGTACATATCTGTCTCAGCTCCGTCCA

ref|AC_000066.1| ATGTGTATATGTACATATCTGTCTCAGCTCCGTCCA

ref|AC_000155.1| ATGTGTATATGTACATATCTGTCTCAGCTCCGTCCA

************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604469	NC_000023	154019738--154019777	20.0	100	80
	AC_000066	154525386--154525429	22.0	100	88
	AC_000155	142911536--142911557	11.0	100	44

New!

WARNING!

This microsatellite marker was firstly predicted In-Silico as informative ^[1], and it has recently been tried-and-tested, apparently (no primer sequences provided), using a pair set of ampliprimers different to the one here provided ^[2]. The authors reported that genotyping of this STR marker yielded peak patterns that made allele calling difficult and, therefore, excluded it form the multiplex assay. Worth of note, the latter study also reported similar impairing effects for two other STR marker loci (REN90371 and REN89898), all of which contain a [AT] repeat unit.

^[1]

^[2]

stSG604469	Information
Amplicon	114-136 bp
Forward ampliprimer	TCATCGTGCTAATGTGCATGT
Reverse ampliprimer	TATGTGGCCTGCTGAGAATG

>ref|NC_000023.9|

TCATCGTGCTAATGTGCATGTGTTACAGCAAGTTACTAAGTTAAATAGAGATTAATTTCATATATATATATATATATATATATATATATA

TATATATATGTTTGCAAAGCATCATTCTCAGCAGGCCACATA

>ref|AC_000066.1|

TCATCGTGCTAATGTGCATGTGTTACAGCAAGTTACTAAGTTAAATAGAGATTAATTTCATATATATATATATATATATATATATATATA

TATATATATATATGTTTGCAAAGCATCATTCTCAGCAGGCCACATA

>ref|AC_000155.1|

TCATCGTGCTAATGTGCATGTGTTACAGCAAGTTACTAAGTTAAATAGAGATTAATTTCATATATATATATATATATATATGTTTGCAAA

GCATCATTCTCAGCAGGCCACATA

stSG604469

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 132 bp

Sequence 2: ref|AC_000066.1| 136 bp

Sequence 3: ref|AC_000155.1| 114 bp

ref|AC_000066.1| TCATCGTGCTAATGTGCATGTGTTACAGCAAGTTACTAAGTTAAATAGAGATTAATTTCA

ref|AC_000155.1| TCATCGTGCTAATGTGCATGTGTTACAGCAAGTTACTAAGTTAAATAGAGATTAATTTCA

ref|NC_000023.9| TCATCGTGCTAATGTGCATGTGTTACAGCAAGTTACTAAGTTAAATAGAGATTAATTTCA

************************************************************

ref|AC_000066.1| TATATATATATATATATATATATATATATATATATATATATATGTTTGCAAAGCATCATT

ref|AC_000155.1| TATATATATATATATATATA----------------------TGTTTGCAAAGCATCATT

ref|NC_000023.9| TATATATATATATATATATATATATATATATATATATA----TGTTTGCAAAGCATCATT

******************** ******************

ref|AC_000066.1| CTCAGCAGGCCACATA

ref|AC_000155.1| CTCAGCAGGCCACATA

ref|NC_000023.9| CTCAGCAGGCCACATA

****************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
stSG604486	NC_000023	154050935--154050987	26.5	100	106
	AC_000066	154556080--154556120	20.5	100	82
	AC_000155	142942378--142942418	20.5	100	82

*New!*	This microsatellite marker has been recently validated experimentally as informative, yet under a different coined name by Ding et al. (2012)^[1]. *WARNING*: We recently warned about the need to acknowledge accurate tracking genetic markers in haemophilia linkage analysis to avoid unnecessary priority claims, as in this case ^[2].
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[3]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.59 and 0.51, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Ding QL, Lu YL, Dai J, Xi XD, Wan XF, Wang HL. Characterisation and validation of a novel panel of the six short tandem repeats for genetic counselling in Chinese haemophilia A pedigrees. Haemophilia 2012, 18(4):621-625
^[2]	Machado FB, Medina-Acosta E (2012) Accurate tracking genetic markers in haemophilia linkage analysis: a matter beyond scientific priority. Haemophilia 18:e359--e360.
^[3]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

stSG604486	Information
Amplicon	204-216 bp
Forward ampliprimer	TGGCAATTTCAGATCACTTCTTT
Reverse ampliprimer	GGTTTTCTTTGTGGCCAAGT

>ref|NC_000023.9|

TGGCAATTTCAGATCACTTCTTTCATTATTAGACAGGTCTAGTAGATAAAACATAAGTAAAGACGAAGGAATTCGAATAAAACATAATAA

ATAAATAACAATAATAAACCAGAACACACACACACACACACACACACACACACACACACACACACACACACACACAGAACGTTCATTTTC

TTTAAAGCAATAATATACTTGGCCACAAAGAAAACC

>ref|AC_000066.1|

TGGCAATTTCAGATCACTTCTTTCATTATTAGACAGGTCTAGTAGATAAAACATAAGTAAAGACGAAGGAATTCGAATAAAACATAATAA

ATAAATAACAATAATAAACCAGAACACACACACACACACACACACACACACACACACACACACAGAACGTTCATTTTCTTTAAAGCAATA

ATATACTTGGCCACAAAGAAAACC

>ref|AC_000155.1|

TGGCAATTTCAGATCACTTCTTTCATTATTAGACAGGTCTAGTAGATAAAACATAAGTAAAGACGAAGGAATTCGAATAAAACATAATAA

ATAAATAACAATAATAAACCAGAACACACACACACACACACACACACACACACACACACACACAGAACGTTCATTTTCTTTAAAGCAATA

ATATACTTGGCCACAAAGAAAACC

stSG604486

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 216 bp

Sequence 2: ref|AC_000066.1| 204 bp

Sequence 3: ref|AC_000155.1| 204 bp

ref|AC_000066.1| TGGCAATTTCAGATCACTTCTTTCATTATTAGACAGGTCTAGTAGATAAAACATAAGTAA

ref|AC_000155.1| TGGCAATTTCAGATCACTTCTTTCATTATTAGACAGGTCTAGTAGATAAAACATAAGTAA

ref|NC_000023.9| TGGCAATTTCAGATCACTTCTTTCATTATTAGACAGGTCTAGTAGATAAAACATAAGTAA

************************************************************

ref|AC_000066.1| AGACGAAGGAATTCGAATAAAACATAATAAATAAATAACAATAATAAACCAGAACACACA

ref|AC_000155.1| AGACGAAGGAATTCGAATAAAACATAATAAATAAATAACAATAATAAACCAGAACACACA

ref|NC_000023.9| AGACGAAGGAATTCGAATAAAACATAATAAATAAATAACAATAATAAACCAGAACACACA

************************************************************

ref|AC_000066.1| CACACACACACACACACACACACACACACACACA------------GAACGTTCATTTTC

ref|AC_000155.1| CACACACACACACACACACACACACACACACACA------------GAACGTTCATTTTC

ref|NC_000023.9| CACACACACACACACACACACACACACACACACACACACACACACAGAACGTTCATTTTC

********************************** **************

ref|AC_000066.1| TTTAAAGCAATAATATACTTGGCCACAAAGAAAACC

ref|AC_000155.1| TTTAAAGCAATAATATACTTGGCCACAAAGAAAACC

ref|NC_000023.9| TTTAAAGCAATAATATACTTGGCCACAAAGAAAACC

************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**VBP1Int4**	NC_000023	154116577--154116620	22.0	95	79
	AC_000066	154621861--154621904	22.0	95	79
	AC_000155	143007387--143007430	22.0	95	79

VBP1Int4	Information
Amplicon	183 bp
Forward ampliprimer	GGTCAGTTTCCCCCAGGTAT
Reverse ampliprimer	CAAAGCAGGGGACCAAAAA

>ref|NC_000023.9|

GGTCAGTTTCCCCCAGGTATACAAAGTACCCTTTCATTTCATTTTGCGTGTCATTTCACCCAGTGTGTGTGTGTGTATGTGTGTGTGTGT

GTGTGTGTGTGTGTGTAGTATGTGCATGTGTCACATTTCAGGAACATTTTGTTGAAATGAAGGATTTAGAAATTTTTTTGGTCCCCTGCT

TTG

>ref|AC_000066.1|

GGTCAGTTTCCCCCAGGTATACAAAGTACCCTTTCATTTCATTTTGCGTGTCATTTCACCCAGTGTGTGTGTGTGTATGTGTGTGTGTGT

GTGTGTGTGTGTGTGTAGTATGTGCATGTGTCACATTTCAGGAACATTTTGTTGAAATGAAGGATTTAGAAATTTTTTTGGTCCCCTGCT

TTG

>ref|AC_000155.1|

GGTCAGTTTCCCCCAGGTATACAAAGTACCCTTTCATTTCATTTTGCGTGTCATTTCACCCAGTGTGTGTGTGTGTATGTGTGTGTGTGT

GTGTGTGTGTGTGTGTAGTATGTGCATGTGTCACATTTCAGGAACATTTTGTTGAAATGAAGGATTTAGAAATTTTTTTGGTCCCCTGCT

TTG

VBP1Int4

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 183 bp

Sequence 2: ref|AC_000066.1| 183 bp

Sequence 3: ref|AC_000155.1| 183 bp

ref|AC_000066.1| GGTCAGTTTCCCCCAGGTATACAAAGTACCCTTTCATTTCATTTTGCGTGTCATTTCACC

ref|NC_000023.9| GGTCAGTTTCCCCCAGGTATACAAAGTACCCTTTCATTTCATTTTGCGTGTCATTTCACC

ref|AC_000155.1| GGTCAGTTTCCCCCAGGTATACAAAGTACCCTTTCATTTCATTTTGCGTGTCATTTCACC

************************************************************

ref|AC_000066.1| CAGTGTGTGTGTGTGTATGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGTATGTGCATGTG

ref|NC_000023.9| CAGTGTGTGTGTGTGTATGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGTATGTGCATGTG

ref|AC_000155.1| CAGTGTGTGTGTGTGTATGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGTATGTGCATGTG

************************************************************

ref|AC_000066.1| TCACATTTCAGGAACATTTTGTTGAAATGAAGGATTTAGAAATTTTTTTGGTCCCCTGCT

ref|NC_000023.9| TCACATTTCAGGAACATTTTGTTGAAATGAAGGATTTAGAAATTTTTTTGGTCCCCTGCT

ref|AC_000155.1| TCACATTTCAGGAACATTTTGTTGAAATGAAGGATTTAGAAATTTTTTTGGTCCCCTGCT

************************************************************

ref|AC_000066.1| TTG

ref|NC_000023.9| TTG

ref|AC_000155.1| TTG

***

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154126.0	NC_000023	154126085--154126111	13.5	100	54
	AC_000066	154631369--154631394	13.5	92	45
	AC_000155	143016895--143016921	13.5	100	54

HEMA154126.0	Information
Amplicon	274-275 bp
Forward ampliprimer	TTTCACTGCCTTGGTTAAATG
Reverse ampliprimer	CAGAGGGTACATGTGCAGGTT

>ref|NC_000023.9|

TTTCACTGCCTTGGTTAAATGTATTCCTAGGTATTTAAATTCTAATTTCATAAAAAATTAGATTTTTAACTAATTTTCAACTTTACTATA

GCAACTGGTTGCAATACCAGAGTAAAATGGCAATGCTGGCTTTGTGTGTGTGTGTGTGTGTGTGTGTGTCACACTCAGTGTGTCACACTG

AGACACTGATACCAGTGATGCATTAAAAAGTATGGTGCACAGACGTCACCACTATGCAATATATTTATGTAACAAACCTGCACATGTACC

CTCTG

>ref|AC_000066.1|

TTTCACTGCCTTGGTTAAATGTATTCCTAGGTATTTAAATTCTAATTTCATAAAAAATTAGATTTTTAACTAATTTTCAACTTTACTATA

GCAACTGGTTGCAATACCAGAGTAAAATGGCAATGCTGGCTTTGTGTGTGTGTGTGTGTGTTGTGTGTCACACTCAGTGTGTCACACTGA

GACACTGATACCAGTGATGCATTAAAAAGTATGGTGCACAGACGTCACCACTATGCAATATATTTATGTAACAAACCTGCACATGTACCC

TCTG

>ref|AC_000155.1|

TTTCACTGCCTTGGTTAAATGTATTCCTAGGTATTTAAATTCTAATTTCATAAAAAATTAGATTTTTAACTAATTTTCAACTTTACTATA

GCAACTGGTTGCAATACCAGAGTAAAATGGCAATGCTGGCTTTGTGTGTGTGTGTGTGTGTGTGTGTGTCACACTCAGTGTGTCACACTG

AGACACTGATACCAGTGATGCATTAAAAAGTATGGTGCACAGACGTCACCACTATGCAATATATTTATGTAACAAACCTGCACATGTACC

CTCTG

HEMA154126.0

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 275 bp

Sequence 2: ref|AC_000066.1| 274 bp

Sequence 3: ref|AC_000155.1| 275 bp

ref|NC_000023.9| TTTCACTGCCTTGGTTAAATGTATTCCTAGGTATTTAAATTCTAATTTCATAAAAAATTA

ref|AC_000155.1| TTTCACTGCCTTGGTTAAATGTATTCCTAGGTATTTAAATTCTAATTTCATAAAAAATTA

ref|AC_000066.1| TTTCACTGCCTTGGTTAAATGTATTCCTAGGTATTTAAATTCTAATTTCATAAAAAATTA

************************************************************

ref|NC_000023.9| GATTTTTAACTAATTTTCAACTTTACTATAGCAACTGGTTGCAATACCAGAGTAAAATGG

ref|AC_000155.1| GATTTTTAACTAATTTTCAACTTTACTATAGCAACTGGTTGCAATACCAGAGTAAAATGG

ref|AC_000066.1| GATTTTTAACTAATTTTCAACTTTACTATAGCAACTGGTTGCAATACCAGAGTAAAATGG

************************************************************

ref|NC_000023.9| CAATGCTGGCTTTGTGTGTGTGTGTGTGTGTGTGTGTGTCACACTCAGTGTGTCACACTG

ref|AC_000155.1| CAATGCTGGCTTTGTGTGTGTGTGTGTGTGTGTGTGTGTCACACTCAGTGTGTCACACTG

ref|AC_000066.1| CAATGCTGGCTTTGTGTGTGTGTGTGTGTGT-TGTGTGTCACACTCAGTGTGTCACACTG

******************************* ****************************

ref|NC_000023.9| AGACACTGATACCAGTGATGCATTAAAAAGTATGGTGCACAGACGTCACCACTATGCAAT

ref|AC_000155.1| AGACACTGATACCAGTGATGCATTAAAAAGTATGGTGCACAGACGTCACCACTATGCAAT

ref|AC_000066.1| AGACACTGATACCAGTGATGCATTAAAAAGTATGGTGCACAGACGTCACCACTATGCAAT

************************************************************

ref|NC_000023.9| ATATTTATGTAACAAACCTGCACATGTACCCTCTG

ref|AC_000155.1| ATATTTATGTAACAAACCTGCACATGTACCCTCTG

ref|AC_000066.1| ATATTTATGTAACAAACCTGCACATGTACCCTCTG

***********************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154130.5	NC_000023	154130572--154130609	19.0	100	76
	AC_000066	154635855--154635892	19.0	100	76
	AC_000155	143021339--143021376	19.0	100	76

*New!*	This microsatellite marker has been recently validated experimentally as informative, yet under a different coined name by Ding et al. (2012)^[1]. *WARNING!* We recently warrant about the need to acknowledge accurate tracking genetic markers in haemophilia linkage analysis to avoid unnecessary priority claims, as in this case ^[2].
New!	This locus has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[3]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.77 and 0.59, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Ding QL, Lu YL, Dai J, Xi XD, Wan XF, Wang HL. Characterisation and validation of a novel panel of the six short tandem repeats for genetic counselling in Chinese haemophilia A pedigrees. Haemophilia 2012, 18(4):621-625
^[2]	Machado FB, Medina-Acosta E (2012) Accurate tracking genetic markers in haemophilia linkage analysis: a matter beyond scientific priority. Haemophilia 18:e359--e360.
^[3]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

HEMA154130.5	Information
Amplicon	162 bp
Forward ampliprimer	ATGATGTACAAGCCCTACCTGA
Reverse ampliprimer	TGAGCCAGGGAAGAATGAAA

>ref|NC_000023.9|

ATGATGTACAAGCCCTACCTGATCTGGGTCCCACACCCCCTCTCTAAATTAATCTATGTGAGTCAGCCCCTGCCCCCCCCCACCCCGCTC

TCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTTCATTTACCCCCCTTTCATTCTTCCCTGGCTCA

>ref|AC_000066.1|

ATGATGTACAAGCCCTACCTGATCTGGGTCCCACACCCCCTCTCTAAATTAATCTATGTGAGTCAGCCCCTGCCCCCCCCCACCCCGCTC

TCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTTCATTTACCCCCCTTTCATTCTTCCCTGGCTCA

>ref|AC_000155.1|

ATGATGTACAAGCCCTACCTGATCTGGGTCCCACACCCCCTCTCTAAATTAATCTATGTGAGTCAGCCCCTGCCCCCCCCCACCCCGCTC

TCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTTTCTTCATTTACCCCCCTTTCATTCTTCCCTGGCTCA

HEMA154130.5

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 162 bp

Sequence 2: ref|AC_000066.1| 162 bp

Sequence 3: ref|AC_000155.1| 162 bp

ref|NC_000023.9| ATGATGTACAAGCCCTACCTGATCTGGGTCCCACACCCCCTCTCTAAATTAATCTATGTG

ref|AC_000155.1| ATGATGTACAAGCCCTACCTGATCTGGGTCCCACACCCCCTCTCTAAATTAATCTATGTG

ref|AC_000066.1| ATGATGTACAAGCCCTACCTGATCTGGGTCCCACACCCCCTCTCTAAATTAATCTATGTG

************************************************************

ref|NC_000023.9| AGTCAGCCCCTGCCCCCCCCCACCCCGCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTC

ref|AC_000155.1| AGTCAGCCCCTGCCCCCCCCCACCCCGCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTC

ref|AC_000066.1| AGTCAGCCCCTGCCCCCCCCCACCCCGCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTC

************************************************************

ref|NC_000023.9| TCTCTTTCTTCATTTACCCCCCTTTCATTCTTCCCTGGCTCA

ref|AC_000155.1| TCTCTTTCTTCATTTACCCCCCTTTCATTCTTCCCTGGCTCA

ref|AC_000066.1| TCTCTTTCTTCATTTACCCCCCTTTCATTCTTCCCTGGCTCA

******************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**CLIC2Int1**	NC_000023	154208272--154208313	21.0	100	84
	AC_000066	154713571--154713612	21.0	100	84
	AC_000155	143098838--143098883	23.0	100	92

CLIC2Int1	Information
Amplicon	173-177 bp
Forward ampliprimer	GCACATGCATCCAAGAACTG
Reverse ampliprimer	CTCCAATTCCTTCCAGGTTG

>ref|NC_000023.9|

GCACATGCATCCAAGAACTGAAAGTATAATAAAAAAGAAAATGTGATATATATATATATATATATATATATATATATATATATATATGCA

CACATACACACACCATGGAATACTACTCAGCCATAAAAAGGCACTAAATAATGGCATTTGCAGCAACCTGGAAGGAATTGGAG

>ref|AC_000066.1|

GCACATGCATCCAAGAACTGAAAGTATAATAAAAAAGAAAATGTGATATATATATATATATATATATATATATATATATATATATATGCA

CACATACACACACCATGGAATACTACTCAGCCATAAAAAGGCACTAAATAATGGCATTTGCAGCAACCTGGAAGGAATTGGAG

>ref|AC_000155.1|

GCACATGCATCCAAGAACTGAAAGTATAATAAAAAAGAAAATGTGATATATATATATATATATATATATATATATATATATATATATATA

TGCACACATACACACACCATGGAATACTACTCAGCCATAAAAAGGCACTAAATAATGGCATTTGCAGCAACCTGGAAGGAATTGGAG

CLIC2Int1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 173 bp

Sequence 2: ref|AC_000066.1| 173 bp

Sequence 3: ref|AC_000155.1| 177 bp

ref|NC_000023.9| GCACATGCATCCAAGAACTGAAAGTATAATAAAAAAGAAAATGTGATATATATATATATA

ref|AC_000066.1| GCACATGCATCCAAGAACTGAAAGTATAATAAAAAAGAAAATGTGATATATATATATATA

ref|AC_000155.1| GCACATGCATCCAAGAACTGAAAGTATAATAAAAAAGAAAATGTGATATATATATATATA

************************************************************

ref|NC_000023.9| TATATATATATATATATATATATATAT----GCACACATACACACACCATGGAATACTAC

ref|AC_000066.1| TATATATATATATATATATATATATAT----GCACACATACACACACCATGGAATACTAC

ref|AC_000155.1| TATATATATATATATATATATATATATATATGCACACATACACACACCATGGAATACTAC

*************************** *****************************

ref|NC_000023.9| TCAGCCATAAAAAGGCACTAAATAATGGCATTTGCAGCAACCTGGAAGGAATTGGAG

ref|AC_000066.1| TCAGCCATAAAAAGGCACTAAATAATGGCATTTGCAGCAACCTGGAAGGAATTGGAG

ref|AC_000155.1| TCAGCCATAAAAAGGCACTAAATAATGGCATTTGCAGCAACCTGGAAGGAATTGGAG

*********************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154229.0	NC_000023	154229069--154229113	11.8	95	76
	AC_000066	154734321--154734361	10.8	94	68
	AC_000155	143119641--143119681	10.8	94	68

*WARNING!*
The published HEMA154229.0 primer pair sequences^[1] anneal to more than one location on contig(s) by e-PCR: chrX:154228869+154229259 and chrX:154377350-154377732 on human chromosome X. The locations are identical at each side of the STR element in over 150 nucleotides. Therefore, the marker HEMA154229.0 cannot be considered unique (UniSTS). (Updated September 12, 2008).
^[1]	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.

HEMA154229.0	Information
Amplicon	96-100 bp
Forward ampliprimer	AAGCCTGGGAGGATGGACTA
Reverse ampliprimer	GGGCGACAGAGCGAGACT

>ref|NC_000023.9|

AAGCCTGGGAGGATGGACTATTATATTCTTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTCTTTGAGACAGAGTCTCGC

TCTGTCGCCC

>ref|AC_000066.1|

AAGCCTGGGAGGATGGACTATTATATTCTTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTCTTTGAGACAGAGTCTCGCTCTG

TCGCCC

>ref|AC_000155.1|

AAGCCTGGGAGGATGGACTATTATATTCTTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTCTTTGAGACAGAGTCTCGCTCTG

TCGCCC

HEMA154229.0

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 100 bp

Sequence 2: ref|AC_000066.1| 96 bp

Sequence 3: ref|AC_000155.1| 96 bp

ref|AC_000066.1| AAGCCTGGGAGGATGGACTATTATATTCTTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

ref|AC_000155.1| AAGCCTGGGAGGATGGACTATTATATTCTTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

ref|NC_000023.9| AAGCCTGGGAGGATGGACTATTATATTCTTTTTCTTTCTTTCTTTCTTTCTTTCTTTCTT

************************************************************

ref|AC_000066.1| T----TTTTTTCTTTGAGACAGAGTCTCGCTCTGTCGCCC

ref|AC_000155.1| T----TTTTTTCTTTGAGACAGAGTCTCGCTCTGTCGCCC

ref|NC_000023.9| TCTTTTTTTTTCTTTGAGACAGAGTCTCGCTCTGTCGCCC

* ***********************************

>chrX:154228869+154229259 391bp AGGAGGTTAATAACACCTTT GCCAGGCGCGGTGGCACGCGCCTG

AGGAGGTTAATAACACCTTTccttctgggatgatgaaataagtaaattaa

ataatattttcacaaatgcctaacccagcactagaagtttctggaaatga

caatgggtgtcaacagcctggtgaagactgtgcttcctcactccattctc

cctccaggcatggctccttgaagcctgggaggatggactattatattctt

tttctttctttctttctttctttctttctttctttttttttctttgagac

agagtctcgctctgtcgcccaggctggagtgcagtggcgcgatcttggct

cactgcagcgtccgccacccaggttcaagtgattctcctgcctcagtctc

ccaagtagctgggactaCAGGCGCGTGCCACCGCGCCTGGC

>chrX:154377350-154377732 383bp AGGAGGTTAATAACACCTTT GCCAGGCGCGGTGGCACGCGCCTG

AGGAGGTTAATAACACCTTTccttctgggatgatgaaataagtaaattaa

ataatattttcacaaatgcctaacccagcactagaagtttctggaaatga

caatgggtgtcaacagcctggtgaagactgtgcttcctcactccattctc

cctccaggcatggctccttgaagcctgggaggatggactattatattctt

tttctttctttctttctttctttttttttttttctttgagacagagtctc

gctctgtcgcccaggctggagtgcagtggcgcgatcttggctcactgcag

cgtccgccacccaggttcaagtgattctcctgcctcagtctcccaagtag

ctgggactaCAGGCGCGTGCCACCGCGCCTGGC

Sequence 1: chrX_154228869+154229259_391bp 391 bp

Sequence 2: chrX_154377350-154377732_383bp 383 bp

chrX_154228869+154229259_391bp AGGAGGTTAATAACACCTTTCCTTCTGGGATGATGAAATAAGTAAATTAA

chrX_154377350-154377732_383bp AGGAGGTTAATAACACCTTTCCTTCTGGGATGATGAAATAAGTAAATTAA

**************************************************

chrX_154228869+154229259_391bp ATAATATTTTCACAAATGCCTAACCCAGCACTAGAAGTTTCTGGAAATGA

chrX_154377350-154377732_383bp ATAATATTTTCACAAATGCCTAACCCAGCACTAGAAGTTTCTGGAAATGA

**************************************************

chrX_154228869+154229259_391bp CAATGGGTGTCAACAGCCTGGTGAAGACTGTGCTTCCTCACTCCATTCTC

chrX_154377350-154377732_383bp CAATGGGTGTCAACAGCCTGGTGAAGACTGTGCTTCCTCACTCCATTCTC

**************************************************

chrX_154228869+154229259_391bp CCTCCAGGCATGGCTCCTTGAAGCCTGGGAGGATGGACTATTATATTCTT

chrX_154377350-154377732_383bp CCTCCAGGCATGGCTCCTTGAAGCCTGGGAGGATGGACTATTATATTCTT

**************************************************

chrX_154228869+154229259_391bp TTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTCTTTTTTTTTCTTTGAGAC

chrX_154377350-154377732_383bp TTTCTTTCTTTCTTTCTTTCTTT--------TTTTTTTTTTCTTTGAGAC

*********************** ******************

chrX_154228869+154229259_391bp AGAGTCTCGCTCTGTCGCCCAGGCTGGAGTGCAGTGGCGCGATCTTGGCT

chrX_154377350-154377732_383bp AGAGTCTCGCTCTGTCGCCCAGGCTGGAGTGCAGTGGCGCGATCTTGGCT

**************************************************

chrX_154228869+154229259_391bp CACTGCAGCGTCCGCCACCCAGGTTCAAGTGATTCTCCTGCCTCAGTCTC

chrX_154377350-154377732_383bp CACTGCAGCGTCCGCCACCCAGGTTCAAGTGATTCTCCTGCCTCAGTCTC

**************************************************

chrX_154228869+154229259_391bp CCAAGTAGCTGGGACTACAGGCGCGTGCCACCGCGCCTGGC

chrX_154377350-154377732_383bp CCAAGTAGCTGGGACTACAGGCGCGTGCCACCGCGCCTGGC

*****************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154279.5(HA544)	NC_000023	154279540--154279610	23.7	100	142
	AC_000066	154785848--154785882	11.7	100	70
	AC_000066	154785881--154785909	9.7	100	58
	AC_000155	143170838--143170872	11.7	100	70
	AC_000155	143170871--143170902	10.7	100	64

HEMA154279.5(HA544)	Information
Amplicon	157-175 bp
Forward ampliprimer	AAAGTGAAACTGTCCGTGTTTG
Reverse ampliprimer	TTGCAGTGAGGTGAGATTGC

>ref|NC_000023.9|

AAAGTGAAACTGTCCGTGTTTGTTAATTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTC

TTCTTCTTATTATTATTATTTTGAGATGGAGTCTCACTCTGTCACCCAGGCTGGAGTGCAGTGGTGCAATCTCACCTCACTGCAA

>ref|AC_000155.1|

AAAGTGAAACTGTCCGTGTTTGTTAATTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTATTATTATTATTATTATTATTATTATTA

TTTTGAGATGGAGTCTCACTCTGTCACCCAGGCTGGAGTGCAGTGGTGCAATCTCACCTCACTGCAA

HEMA154279.5(HA544)

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 175 bp

Sequence 3: ref|AC_000155.1| 157 bp

ref|NC_000023.9| AAAGTGAAACTGTCCGTGTTTGTTAATTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTC

ref|AC_000155.1| AAAGTGAAACTGTCCGTGTTTGTTAATTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTC

************************************************************

ref|NC_000023.9| TTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTCTTATTATTATTATTTTGAGATGGA

ref|AC_000155.1| TTATTATTA------------------TTATTATTATTATTATTATTATTTTGAGATGGA

** ** ** ** ** ** ************************

ref|NC_000023.9| GTCTCACTCTGTCACCCAGGCTGGAGTGCAGTGGTGCAATCTCACCTCACTGCAA

ref|AC_000155.1| GTCTCACTCTGTCACCCAGGCTGGAGTGCAGTGGTGCAATCTCACCTCACTGCAA

*******************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154281.2	NC_000023	154281204--154281236	16.5	93	57
	AC_000066	154787342--154787374	16.5	93	57
	AC_000155	143172484--143172516	16.5	93	57

HEMA154281.2	Information
Amplicon	378 bp
Forward ampliprimer	TCTGCACTCCCATGTTCATT
Reverse ampliprimer	CTGTCCACTCCCCTACTCCA

>ref|NC_000023.9|

TCTGCACTCCCATGTTCATTGCAGCACTATCACAATAGCCAAGATATGAAATAAACCTGTTTCCATCAACAGATGTAATGGATTTTTAAAAT

GTGGTGTGTGTATACATACACACACACACACACACACGCACACACACACATATAGCTATATACACACACATGCACACAATGTAATATTATTC

AACCTTAACAGAAATTTTTGCCATTTGCAACAACACAGATAAACTTAGAGGACATTATGCTAAGTGAAATAAACCAGGCACAGAGAGACAAA

TATTACATTATCTGACTTATATGTGCAATCTAAAAAAGTCAAATTTATAGAAGTAAAGAGTAGAATGGTAGTTGCCAGGGGCTGGAGTAGGG

GAGTGGACAG

>ref|AC_000066.1|

TCTGCACTCCCATGTTCATTGCAGCACTATCACAATAGCCAAGATATGAAATAAACCTGTTTCCATCAACAGATGTAATGGATTTTTAAAAT

GTGGTGTGTGTATGTATACACACACACACACACACACGCACACACACACATATAGCTATATACACACACATGCACACAATGTAATATTATTC

AACCTTAACAGAAATTTTTGCCATTTGCAACAACACAGATAAACTTAGAGGACATTATGCTAAGTGAAATAAACCAGGCACAGAGAGACAAA

TATTACATTATCTGACTTATATGTGCAATCTAAAAAAGTCAAATTTATAGAAGTAAAGAGTAGAATGGTAGTTGCCAGGGGCTGGAGTAGGG

GAGTGGACAG

>ref|AC_000155.1|

TCTGCACTCCCATGTTCATTGCAGCACTATCACAATAGCCAAGATATGAAATAAACCTGTTTCCATCAACAGATGTAATGGATTTTTAAAAT

GTGGTGTGTGTATGTATACACACACACACACACACACGCACACACACACATATAGCTATATACACACACATGCACACAATGTAATATTATTC

AACCTTAACAGAAATTTTTGCCATTTGCAACAACACAGATAAACTTAGAGGACATTATGCTAAGTGAAATAAACCAGGCACAGAGAGACAAA

TATTACATTATCTGACTTATATGTGCAATCTAAAAAAGTCAAATTTATAGAAGTAAAGAGTAGAATGGTAGTTGCCAGGGGCTGGAGTAGGG

GAGTGGACAG

HEMA154281.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 378 bp

Sequence 2: ref|AC_000066.1| 378 bp

Sequence 3: ref|AC_000155.1| 378 bp

ref|NC_000023.9| TCTGCACTCCCATGTTCATTGCAGCACTATCACAATAGCCAAGATATGAAATAAACCTGT

ref|AC_000155.1| TCTGCACTCCCATGTTCATTGCAGCACTATCACAATAGCCAAGATATGAAATAAACCTGT

ref|AC_000066.1| TCTGCACTCCCATGTTCATTGCAGCACTATCACAATAGCCAAGATATGAAATAAACCTGT

************************************************************

ref|NC_000023.9| TTCCATCAACAGATGTAATGGATTTTTAAAATGTGGTGTGTGTATACATACACACACACA

ref|AC_000155.1| TTCCATCAACAGATGTAATGGATTTTTAAAATGTGGTGTGTGTATGTATACACACACACA

ref|AC_000066.1| TTCCATCAACAGATGTAATGGATTTTTAAAATGTGGTGTGTGTATGTATACACACACACA

********************************************* *************

ref|NC_000023.9| CACACACACGCACACACACACATATAGCTATATACACACACATGCACACAATGTAATATT

ref|AC_000155.1| CACACACACGCACACACACACATATAGCTATATACACACACATGCACACAATGTAATATT

ref|AC_000066.1| CACACACACGCACACACACACATATAGCTATATACACACACATGCACACAATGTAATATT

************************************************************

ref|NC_000023.9| ATTCAACCTTAACAGAAATTTTTGCCATTTGCAACAACACAGATAAACTTAGAGGACATT

ref|AC_000155.1| ATTCAACCTTAACAGAAATTTTTGCCATTTGCAACAACACAGATAAACTTAGAGGACATT

ref|AC_000066.1| ATTCAACCTTAACAGAAATTTTTGCCATTTGCAACAACACAGATAAACTTAGAGGACATT

************************************************************

ref|NC_000023.9| ATGCTAAGTGAAATAAACCAGGCACAGAGAGACAAATATTACATTATCTGACTTATATGT

ref|AC_000155.1| ATGCTAAGTGAAATAAACCAGGCACAGAGAGACAAATATTACATTATCTGACTTATATGT

ref|AC_000066.1| ATGCTAAGTGAAATAAACCAGGCACAGAGAGACAAATATTACATTATCTGACTTATATGT

************************************************************

ref|NC_000023.9| GCAATCTAAAAAAGTCAAATTTATAGAAGTAAAGAGTAGAATGGTAGTTGCCAGGGGCTG

ref|AC_000155.1| GCAATCTAAAAAAGTCAAATTTATAGAAGTAAAGAGTAGAATGGTAGTTGCCAGGGGCTG

ref|AC_000066.1| GCAATCTAAAAAAGTCAAATTTATAGAAGTAAAGAGTAGAATGGTAGTTGCCAGGGGCTG

************************************************************

ref|NC_000023.9| GAGTAGGGGAGTGGACAG

ref|AC_000155.1| GAGTAGGGGAGTGGACAG

ref|AC_000066.1| GAGTAGGGGAGTGGACAG

******************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154311.3	NC_000023	154311336--154311361	5.2	100	52
	AC_000066	154817641--154817666	5.2	100	52
	AC_000155	143202733--143202773	8.2	100	82

New!	This microsatellite marker has been recently validated experimentally as informative^[1].
New!	Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[2]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect).
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.
^[2]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.

HEMA154311.3	Information
Amplicon	254-269 bp
Forward ampliprimer	GTTTTGGTCCTGGCACTCAT
Reverse ampliprimer	TTCACGGTTTTAATTTTTCATGTC

>ref|NC_000023.9|

GTTTTGGTCCTGGCACTCATCCATGAGTTTCTTATGAAGAGAAGAGAAGAGAAGAGAAGAGGAGATTACGAAATAAAAATGTGATGTACA

CCTTAATGTGTTATCATTTCCATCAATGAATTTTACAACTTTTCAATAAAATGTTTCTTTTTACTCTGATATCTAGAAGTTCTATTCAAT

GAACACTGAAAAGATATAGACAGATTCATGATAGCTAGATAGATACATTAGACATGAAAAATTAAAACCGTGAA

>ref|AC_000066.1|

GTTTTGGTCCTGGCACTCATCCATGAGTTTCTTATGAAGAGAAGAGAAGAGAAGAGAAGAGGAGATTACGAAATAAAAATGTGATGTACA

CCTTAATGTGTTATCACTTCCATCAATGAATTTTACAACTTTTCAATAAAATGTTTCTTTTTACTCTGATATCTAGAAGTTCTATTCAAT

GAACACTGAAAAGATATAGACAGATTCATGATAGCTAGATAGATAGATTAGACATGAAAAATTAAAACCGTGAA

>ref|AC_000155.1|

GTTTTGGTCCTGGCACTCATCCATGAGTTTCTTATGAAGAGAAGAGAAGAGAAGAGAAGAGAAGAGAAGAGAAGAGGAGATTACGAAATA

AAAATGTGATGTACACCTTAATGTGTTATCACTTCCATCAATGAATTTTACAACTTTTCAATAAAATGTTTCTTTTTACTCTGATATCTA

GAAGTTCTATTCAATGAACACTGAAAAGATATAGACAGATTCATGATAGCTAGATAGATAGATTAGACATGAAAAATTAAAACCGTGAA

HEMA154311.3

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 254 bp

Sequence 2: ref|AC_000066.1| 254 bp

Sequence 3: ref|AC_000155.1| 269 bp

ref|NC_000023.9| GTTTTGGTCCTGGCACTCATCCATGAGTTTCTTATGAAGAGAAGAGAAGAGAAGAGAAGA

ref|AC_000066.1| GTTTTGGTCCTGGCACTCATCCATGAGTTTCTTATGAAGAGAAGAGAAGAGAAGAGAAGA

ref|AC_000155.1| GTTTTGGTCCTGGCACTCATCCATGAGTTTCTTATGAAGAGAAGAGAAGAGAAGAGAAGA

************************************************************

ref|NC_000023.9| G---------------GAGATTACGAAATAAAAATGTGATGTACACCTTAATGTGTTATC

ref|AC_000066.1| G---------------GAGATTACGAAATAAAAATGTGATGTACACCTTAATGTGTTATC

ref|AC_000155.1| GAAGAGAAGAGAAGAGGAGATTACGAAATAAAAATGTGATGTACACCTTAATGTGTTATC

* ********************************************

ref|NC_000023.9| ATTTCCATCAATGAATTTTACAACTTTTCAATAAAATGTTTCTTTTTACTCTGATATCTA

ref|AC_000066.1| ACTTCCATCAATGAATTTTACAACTTTTCAATAAAATGTTTCTTTTTACTCTGATATCTA

ref|AC_000155.1| ACTTCCATCAATGAATTTTACAACTTTTCAATAAAATGTTTCTTTTTACTCTGATATCTA

* **********************************************************

ref|NC_000023.9| GAAGTTCTATTCAATGAACACTGAAAAGATATAGACAGATTCATGATAGCTAGATAGATA

ref|AC_000066.1| GAAGTTCTATTCAATGAACACTGAAAAGATATAGACAGATTCATGATAGCTAGATAGATA

ref|AC_000155.1| GAAGTTCTATTCAATGAACACTGAAAAGATATAGACAGATTCATGATAGCTAGATAGATA

************************************************************

ref|NC_000023.9| CATTAGACATGAAAAATTAAAACCGTGAA

ref|AC_000066.1| GATTAGACATGAAAAATTAAAACCGTGAA

ref|AC_000155.1| GATTAGACATGAAAAATTAAAACCGTGAA

*****************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TMLHEInt5**	NC_000023	154393088--154393119	6.4	100	64
**TMLHEInt5**	AC_000155	143236086--143236127	8.4	100	84

New!	This microsatellite marker has been recently validated experimentally as informative,^[1] albeit using a different (redesigned) pair of primers. *WARNING!* The TMLHEInt5 redesigned reverse primer matches a second location 1918 bp downwards, yielding a 2148 bp TMLHEInt5 specific extended product, and therefore polymorphic due to the STR element, which is excluded from analysis by capillary electrophoresis within the analysis range of 450 bp, and thus it does not interfere with TMLHEInt5 allele calling ranging 219-244 bp.
New!	Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[2]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect).
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.
^[2]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.

TMLHEInt5	Information
Amplicon	286-296 bp
Forward ampliprimer	CCAAAACTAATGGTTAGGCAAT
Reverse ampliprimer	GAGGGATAGCATCGGGAGAT

TMLHEInt5 (redesigned)	Information
Amplicon	219-244
Forward ampliprimer	TCCAAAACTAATGGTTAGGCAAT
Reverse ampliprimer	CACCAGCATGGCACATGTAT

>ref|NC_000023.9|

CCAAAACTAATGGTTAGGCAATACTTTTTTGGGGGATAATTTATAATTACTTTGAAAAATGTAAAATTACATTTTAATAATTAACAAAAT

TAGGACTCCATTTATATAGTAAGATGAGTTTTCTTTTTATTTTATTTTATTTTATTTTATTTTATTATACTTTAAGTTTTAGGGTACATG

TGCACAATGTGCAGGTTTGTTACATATGTATACATGTGCCATGCTGGTGTGCTGCACCCACTAACTCATCATCTAGCATTAGGTATATCT

CCCGATGCTATCCCTC

>ref|AC_000155.1|

CCAAAACTAATGGTTAGGCAATACTTTTTTGGGGGATAATTTATAATTACTTTGAAAAATGTAAAATTACATTTTAATAATTAACAAAATT

AGGACTCCATTTATATAGTAAGATGAGTTTTCTTTTTATTTTATTTTATTTTATTTTATTTTATTTTATTTTATTATACTTTAAGTTTTAG

GGTACATGTGCACAATGTGCAGGTTTGTTACATATGTATACATGTGCCATGCTGGTGTGCTGCACCCACTAACTCATCATCTAGCATTAGG

TATATCTCCCGATGCTATCCCTC

TMLHEInt5 (redesigned) ^[1]validation by iPCR at UCSC

>chrX:154303473+154303702 230bp TCCAAAACTAATGGTTAGGCAAT CACCAGCATGGCACATGTAT

TCCAAAACTAATGGTTAGGCAATacttttttgggggataatttataatta

ctttgaaaaatgtaaaattacattttaataattaacaaaattaggactcc

atttatatagtaagatgagttttctttttattttattttattttatttta

ttttattatactttaagttttagggtacatgtgcacaatgtgcaggtttg

ttacatatgtATACATGTGCCATGCTGGTG

>chrX:154303473+154305620 2148bp TCCAAAACTAATGGTTAGGCAAT CACCAGCATGGCACATGTAT

TCCAAAACTAATGGTTAGGCAATacttttttgggggataatttataatta

ctttgaaaaatgtaaaattacattttaataattaacaaaattaggactcc

atttatatagtaagatgagttttctttttattttattttattttatttta

ttttattatactttaagttttagggtacatgtgcacaatgtgcaggtttg

ttacatatgtatacatgtgccatgctggtgtgctgcacccactaactcat

catctagcattaggtatatctcccgatgctatccctcccccctcccccca

ccccacaacagtccccagagtgtgatgttccccttcctgtgtccatgtgt

tctcattgttcaattcccacctatgagtgagaacatgcggtgtttggttt

gttgttcttgtgatagtttgctgagaatgatggtttccagctccatccat

gtccctacaaaggacatgaactcatcattttttatggctgcatagtattc

catggtgtatatgtgccacattttctttatccagtctatcgttgttggac

atttgggttggttccaagtctttgctattgtgaatagtgccacagtaaac

atacgtgtgcatgtgtctttatagcagcatgatttataatcctttgggta

tatacccagtaatgggatggctgggtcaaatggtatttctagttctagat

ccctgaggaatcgccacactgacttccacaatggttgaactagtttacag

tcccaccaacagtgtaaaagtgttcctatttcttcacatcctctccagca

cctgttgtttcctgactttttaatgattgccattctaactggtgtgagat

ggtatctcattgtggttttgatttgcatttctctgatggccagtaatgat

gagcattttctcatgtgttttttggctgcataaatgtcttcttttgagaa

cttaaaaatattttattcctaggagtctgatatagtttggatatttatcc

ccaccaaaatctcatgttgaattgtaatctccagtgctggaggtagggcc

tggtggaagtgtttagctcatggaggtggataactgatggcttggtgctg

tcttcagatagtgagtgagttcctgtgacagctagctgtttaaaagtgtg

tggcacctcctccctgactctctctcacttgctcctgtttttgccatgtg

aactgtttgctccccgctcaccttccaccatgattggaagctccctgagg

cctccccagaagaagatgctgctatgctttctgtatagcctgcagaacca

tgagacaattaaatctctcttttttttttttataaattacccagtctcag

gtatttatttatagcaatgcaagaatggcctaatacagaatctaaaaact

gagttaaggtatttcaaaatgatcatgtttctaggggaagagaaagtcat

acaaatctgcaggagcagatatgatcttggttcgaacagaatggaaaata

attaagaaaccactcaagactatcacatataaagactttacactgtaaaa

gtctaatccacctacccacagggctcttgaaaataggtagtgtcagtgtg

ccgatccagagctagcttggtgtacgcagtgtcacctctggagaagtctg

aagtgaaataccacatcctcccataaatggtttctctgttagttgaaata

aagaaagacctgttagctatttgaaatgttcattcttgtctttcaaaact

ccttccaataccttctttagttactcacaaataacagtagatttttttgg

ttccagaatttaatccataaggtgaaaatgacagttgaacatgctagtct

tgacaattctgtcacttattaacattctagaaagttggggagaaaactgt

ggaagtataatattgtattatgaaagcttggagaatcatcccactctgat

tatgtgcccagttttcatagccttcataaattcaaggttcttttctatgc

tcacctttctttcacagaaaaagatatagctgcttctggaagtgattttt

ttattttattttattaattattattatactttaagttttagggtacatgt

gcacaatgtgcaggtttgttacatatgtATACATGTGCCATGCTGGTG

TMLHEInt5

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 286 bp

Sequence 2: ref|AC_000155.1| 296 bp

ref|NC_000023.9| CCAAAACTAATGGTTAGGCAATACTTTTTTGGGGGATAATTTATAATTACTTTGAAAAAT

ref|AC_000155.1| CCAAAACTAATGGTTAGGCAATACTTTTTTGGGGGATAATTTATAATTACTTTGAAAAAT

************************************************************

ref|NC_000023.9| GTAAAATTACATTTTAATAATTAACAAAATTAGGACTCCATTTATATAGTAAGATGAGTT

ref|AC_000155.1| GTAAAATTACATTTTAATAATTAACAAAATTAGGACTCCATTTATATAGTAAGATGAGTT

************************************************************

ref|NC_000023.9| TTCTTTTTATTTTATTTTATTTTATTTTATTTTATT----------ATACTTTAAGTTTT

ref|AC_000155.1| TTCTTTTTATTTTATTTTATTTTATTTTATTTTATTTTATTTTATTATACTTTAAGTTTT

************************************ **************

ref|NC_000023.9| AGGGTACATGTGCACAATGTGCAGGTTTGTTACATATGTATACATGTGCCATGCTGGTGT

ref|AC_000155.1| AGGGTACATGTGCACAATGTGCAGGTTTGTTACATATGTATACATGTGCCATGCTGGTGT

************************************************************

ref|NC_000023.9| GCTGCACCCACTAACTCATCATCTAGCATTAGGTATATCTCCCGATGCTATCCCTC

ref|AC_000155.1| GCTGCACCCACTAACTCATCATCTAGCATTAGGTATATCTCCCGATGCTATCCCTC

********************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TMLHEInt2**	NC_000023	154415524--154415567	22.0	100	88
**TMLHEInt2**	AC_000066	154928463--154928508	23.0	100	92

New!	This microsatellite marker was firstly predicted In-Silico as informative ^[1], and it has recently been tried-and-tested under the *THLMEInt2* mis-spelled name (*WARNING!) as informative using a pair set of ampliprimers different to the one here provided ^[2]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.63 and 0.67, respectively, using a tetradecaplex (13 STR loci* plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.
^[2]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

TMLHEInt2	Information
Amplicon	133-136 bp
Forward ampliprimer	TGTCCTAGGGTGTGTGGAAT
Reverse ampliprimer	CACTCTAGGGAAAGGGTTCTCA

>ref|NC_000023.9|

TGTCCTAGGGTGTGTGGAATAATATGCAAAATTCGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAATTTCTATG

GGGTCTATAACCTAAAAATATTGAGAACCCTTTCCCTAGAGTG

>ref|AC_000066.1|

TGTCCTAGGGTGTGTGGAATAATATGCAAAATTCGGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTAGAATTTCT

ATGGGGTCTATAACCTAAAAATATTGAGAACCCTTTCCCTAGAGTG

TMLHEInt2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 133 bp

Sequence 2: ref|AC_000066.1| 136 bp

ref|NC_000023.9| TGTCCTAGGGTGTGTGGAATAATATGCAAAATTCG---TGTGTGTGTGTGTGTGTGTGTG

ref|AC_000066.1| TGTCCTAGGGTGTGTGGAATAATATGCAAAATTCGGTGTGTGTGTGTGTGTGTGTGTGTG

*********************************** **********************

ref|NC_000023.9| TGTGTGTGTGTGTGTGTGTGTAGAATTTCTATGGGGTCTATAACCTAAAAATATTGAGAA

ref|AC_000066.1| TGTGTGTGTGTGTGTGTGTGTAGAATTTCTATGGGGTCTATAACCTAAAAATATTGAGAA

************************************************************

ref|NC_000023.9| CCCTTTCCCTAGAGTG

ref|AC_000066.1| CCCTTTCCCTAGAGTG

****************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TMLHEInt1**.3	NC_000023	154436837--154436875	9.8	100	78
	AC_000066	154949778--154949812	8.8	100	70
	AC_000155	143279663--143279697	8.8	100	70

New!	This microsatellite marker has been recently validated experimentally as informative ^[1].
New!	Multiallelic interlocus non-random association analysis revealed that **GAB3Int1 is at gametic (linkage) EQUILIBRIUM with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1*.1. The observed breakdown of gametic disequilibrium attests historical recombination between GAB3*Int1 and the F8 gene.
^[1]	Machado FB, Alves Da Silva AF, Rossetti LC, De Brasi CD, Medina-Acosta E. Informativeness of a novel multiallelic marker-set comprising an F8 intron 21 and three tightly linked loci for haemophilia A carriership analysis. Haemophilia 2011, 17:257-66.

TMLHEInt1.3	Information
Amplicon	165-169 bp
Forward ampliprimer	ATCTTCCTCCCTCAAGTGCT
Reverse ampliprimer	ATGATCATGCCACTGCACTC

>ref|NC_000023.9|

ATCTTCCTCCCTCAAGTGCTATTATTGTGTATTTTAATTGCATACACATGCACGCCACATGACATTACTATTTTTATTCATTCATTCATT

CATTCATTCATTCATTCATTCATTTATAGATTCAGAGCCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGCATGATCAT

>ref|AC_000066.1|

ATCTTCCTCCCTCAAGTGCTATTATTGTGTATTTTAATTGCATACACATGCACGCCACATGACATTACTATTTTTATTCATTCATTCATT

CATTCATTCATTCATTCATTTATAGATTCAGAGCCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGCATGATCAT

>ref|AC_000155.1|

ATCTTCCTCCCTCAAGTGCTATTATTGTGTATTTTAATTGCATACACATGCACGCCACATGACATTACTATTTTTATTCATTCATTCATT

CATTCATTCATTCATTCATTTATAGATTCAGAGCCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGCATGATCAT

TMLHEInt1.3

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 169 bp

Sequence 2: ref|AC_000066.1| 165 bp

Sequence 3: ref|AC_000155.1| 165 bp

ref|AC_000066.1|      ATCTTCCTCCCTCAAGTGCTATTATTGTGTATTTTAATTGCATACACATGCACGCCACAT

ref|AC_000155.1|      ATCTTCCTCCCTCAAGTGCTATTATTGTGTATTTTAATTGCATACACATGCACGCCACAT

ref|NC_000023.9|      ATCTTCCTCCCTCAAGTGCTATTATTGTGTATTTTAATTGCATACACATGCACGCCACAT

                      ************************************************************

ref|AC_000066.1|      GACATTACTATTTTTATTCATTCATTCATTCATTCATTCATTCATTCATT----TATAGA

ref|AC_000155.1|      GACATTACTATTTTTATTCATTCATTCATTCATTCATTCATTCATTCATT----TATAGA

ref|NC_000023.9|      GACATTACTATTTTTATTCATTCATTCATTCATTCATTCATTCATTCATTCATTTATAGA

                      **************************************************    ******

ref|AC_000066.1|      TTCAGAGCCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGCATGATCAT

ref|AC_000155.1|      TTCAGAGCCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGCATGATCAT

ref|NC_000023.9|      TTCAGAGCCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGCATGATCAT

                      *************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TMLHEInt1**.2	NC_000023	154468260--154468308	24.5	95	89
	AC_000066	154968763--154968807	22.5	95	81
	AC_000155	143298044--143298088	22.5	95	81

TMLHEInt1.2	Information
Amplicon	296-300 bp
Forward ampliprimer	CAAAGGGGGATAATGAAGACA
Reverse ampliprimer	TCTGTGTGCTCCAGGATTGA

>ref|NC_000023.9|

CAAAGGGGGATAATGAAGACATTACATGCAAATGGAAACTAAAAGTAAACAGGAGTAGCTATACTTAGATAAAACAGACTTTAGGTGAAA

AACAGTAAAAAGACAAGGTCATTACATAATGGTAAATTAATCAAGCAAGAGGATATAAATTATACACACACACAAACACACACACACACA

CACACACACACACACACACACATATATATATGCTTTATGAATCTGTGTGCTCCATGAAAATAAACAAAATGTTTTGTATGTTTGGAGTAC

ATATATGCACTCAATCCTGGAGCACACAGA

>ref|AC_000066.1|

CAAAGGGGGATAATGAAGACATTACATGCAAATGGAAACTAAAAGTAAACAGGAGTAGCTATACTTAGATAAAACAGACTTTAGGTGAAA

AACAGTAAAAAGACAAGGTCATTACATAATGGTAAATTAATCAAGCAAGAGGATATAAATTATACACACACACAAACACACACACACACA

CACACACACACACACACATATATATATGCTTTATGAATCTGTGTGCTCCATGAAAATAAACAAAATGTTTTGTATGTTTGGAGTACATAT

ATGCACTCAATCCTGGAGCACACAGA

>ref|AC_000155.1

CAAAGGGGGATAATGAAGACATTACATGCAAATGGAAACTAAAAGTAAACAGGAGTAGCTATACTTAGATAAAACAGACTTTAGGTGAAA

AACAGTAAAAAGACAAGGTCATTACATAATGGTAAATTAATCAAGCAAGAGGATATAAATTATACACACACACAAACACACACACACACA

CACACACACACACACACATATATATATGCTTTATGAATCTGTGTGCTCCATGAAAATAAACAAAATGTTTTGTATGTTTGGAGTACATAT

ATGCACTCAATCCTGGAGCACACAGA

TMLHEInt1.2

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 300 bp

Sequence 2: ref|AC_000066.1| 296 bp

Sequence 3: ref|AC_000155.1| 296 bp

ref|AC_000066.1| CAAAGGGGGATAATGAAGACATTACATGCAAATGGAAACTAAAAGTAAACAGGAGTAGCT

ref|AC_000155.1| CAAAGGGGGATAATGAAGACATTACATGCAAATGGAAACTAAAAGTAAACAGGAGTAGCT

ref|NC_000023.9| CAAAGGGGGATAATGAAGACATTACATGCAAATGGAAACTAAAAGTAAACAGGAGTAGCT

************************************************************

ref|AC_000066.1| ATACTTAGATAAAACAGACTTTAGGTGAAAAACAGTAAAAAGACAAGGTCATTACATAAT

ref|AC_000155.1| ATACTTAGATAAAACAGACTTTAGGTGAAAAACAGTAAAAAGACAAGGTCATTACATAAT

ref|NC_000023.9| ATACTTAGATAAAACAGACTTTAGGTGAAAAACAGTAAAAAGACAAGGTCATTACATAAT

************************************************************

ref|AC_000066.1| GGTAAATTAATCAAGCAAGAGGATATAAATTATACACACACACAAACACACACACACACA

ref|AC_000155.1| GGTAAATTAATCAAGCAAGAGGATATAAATTATACACACACACAAACACACACACACACA

ref|NC_000023.9| GGTAAATTAATCAAGCAAGAGGATATAAATTATACACACACACAAACACACACACACACA

************************************************************

ref|AC_000066.1| CACACACACACACACACA----TATATATATGCTTTATGAATCTGTGTGCTCCATGAAAA

ref|AC_000155.1| CACACACACACACACACA----TATATATATGCTTTATGAATCTGTGTGCTCCATGAAAA

ref|NC_000023.9| CACACACACACACACACACACATATATATATGCTTTATGAATCTGTGTGCTCCATGAAAA

****************** **************************************

ref|AC_000066.1| TAAACAAAATGTTTTGTATGTTTGGAGTACATATATGCACTCAATCCTGGAGCACACAGA

ref|AC_000155.1| TAAACAAAATGTTTTGTATGTTTGGAGTACATATATGCACTCAATCCTGGAGCACACAGA

ref|NC_000023.9| TAAACAAAATGTTTTGTATGTTTGGAGTACATATATGCACTCAATCCTGGAGCACACAGA

************************************************************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
**TMLHEInt1**.1	NC_000023	154486183--154486266	20.5	90	125
**TMLHEInt1**.1	AC_000066	154986681--154986760	19.5	89	108

New!	This microsatellite marker has been recently validated experimentally as informative ^[1].
New!	Multiallelic interlocus non-random association analysis revealed that **GAB3Int1 is at gametic (linkage) EQUILIBRIUM with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1.1. The observed breakdown of gametic disequilibrium attests historical recombination between GAB3Int1** and the F8 gene.
New!	This microsatellite marker was firstly predicted In-Silico as informative ^[1], and it has recently been tried-and-tested under the *THLMEInt1.1* mis-spelled name (*WARNING!) as informative using a pair set of ampliprimers different to the one here provided ^[2]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.61 and 0.57, respectively, using a tetradecaplex (13 STR loci* plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Alves Da Silva AF, Rossetti LC, De Brasi CD, Medina-Acosta E. Informativeness of a novel multiallelic marker-set comprising an F8 intron 21 and three tightly linked loci for haemophilia A carriership analysis. Haemophilia 2011, 17:257-66.
^[2]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

TMLHEInt1.1	Information
Amplicon	247-251 bp
Forward ampliprimer	GTAGTCCCAGGCACTCAGGA
Reverse ampliprimer	CTGGGTTCTTTCCATTTCCA

>ref|NC_000023.9|

GTAGTCCCAGGCACTCAGGAGACTGAGGTGGGAGGATCGTTTGAGCCTGGGAGGTTGAGGATGCAATGAGCTGCAACTGTGTCACTGCAC

TCCAGCCTGAGCAACAGAGCAAGACCCTGTCGAAAGAAAGAAAGAAAAGAAAGAAGAAAAGAAAAGAAAGAATGAAAGAAAGAAAGAAAG

AAAGAAAGAAAGAAAGAAAGAAAGAGTGACAAGATGATCCTAATATTTATATGGAAATGGAAAGAACCCAG

>ref|AC_000066.1|

GTAGTCCCAGGCACTCAGGAGACTGAGGTGGGAGGATCGTTTGAGCCTGGGAGGTTGAGGATGCAATGAGCTGCAACTGTGTCACTGCAC

TCCAGCCTGAGCAACAGAGCAAGACCCTGTCGAAAGAAAGAAAGAAAAGAAAGAAGAAAAGAAAAGAAAGAATGAATGAAAGAAAGAAAG

AAAGAAAGAAAGAAAGAAAGAGTGACAAGATGATCCTAATATTTATATGGAAATGGAAAGAACCCAG

TMLHEInt1.1

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 251 bp

Sequence 2: ref|AC_000066.1| 247 bp

ref|NC_000023.9| GTAGTCCCAGGCACTCAGGAGACTGAGGTGGGAGGATCGTTTGAGCCTGGGAGGTTGAGG

ref|AC_000066.1| GTAGTCCCAGGCACTCAGGAGACTGAGGTGGGAGGATCGTTTGAGCCTGGGAGGTTGAGG

************************************************************

ref|NC_000023.9| ATGCAATGAGCTGCAACTGTGTCACTGCACTCCAGCCTGAGCAACAGAGCAAGACCCTGT

ref|AC_000066.1| ATGCAATGAGCTGCAACTGTGTCACTGCACTCCAGCCTGAGCAACAGAGCAAGACCCTGT

************************************************************

ref|NC_000023.9| CGAAAGAAAGAAAGAAAAGAAAGAAGAAAAGAAAAGAAAGAATGAAAGAAAGAAAGAAAG

ref|AC_000066.1| CGAAAGAAAGAAAGAAAAGAAAGAAGAAAAGAAAAGAAAGAATGAATGAAAGAAAGAAAG

********************************************** *** *********

ref|NC_000023.9| AAAGAAAGAAAGAAAGAAAGAAAGAGTGACAAGATGATCCTAATATTTATATGGAAATGG

ref|AC_000066.1| AAAGAAAGAAAGAAAGAAA----GAGTGACAAGATGATCCTAATATTTATATGGAAATGG

******************* *************************************

ref|NC_000023.9| AAAGAACCCAG

ref|AC_000066.1| AAAGAACCCAG

***********

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154498.9	NC_000023	154498956--154499028	38.0	91	125
	AC_000066	154999445--154999539	49.0	91	160
	AC_000155	143326851--143326945	49.0	91	160

New!	This microsatellite marker was firstly predicted In-Silico as informative ^[1], and it has recently been tried-and-tested as informative using a pair set of ampliprimers different to the one here provided ^[2]. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates of 0.84 and 0.78, respectively, using a tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay, based on the information about the STR loci reported here. The tetradecaplex assay was valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in HEMA at risk couples.
^[1]	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.
^[2]	Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

HEMA154498.9	Information
Amplicon	179-201 bp
Forward ampliprimer	GGATTCCAGTGACATTGACTCTA
Reverse ampliprimer	GAGAGGAGTGGGGAGGAAAG

>ref|NC_000023.9|

GGATTCCAGTGACATTGACTCTATCATTCTTTTACCCCTATTTTTCCCTGCACATTTATCTGGGGTCTCTCTCTCTCTCTCTCTCTCTCT

CTCTCCTCTCTCTCTCTCTCTCTCTTCTCTCTCTCTTCTCTCTCTCTCATCCCCTCGTTCCCTTCCTTGCTTTCCTCCCCACTCCTCTC

>ref|AC_000066.1|

GGATTCCAGTGACATTGACTCTATCATTCTTTTACCCCTATTCTTCCCTGCACATTTATCTGGGGTCTCTCTCCCTCTCTCTCTCTCTCT

CTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCCTCTCTCTCTCTCTCTTCTCTCTCTCTTCTCTCTCTCTCATCCCCTCGTTCCCTTCCTT

GCTTTCCTCCCCACTCCTCTC

>ref|AC_000155.1|

GGATTCCAGTGACATTGACTCTATCATTCTTTTACCCCTATTCTTCCCTGCACATTTATCTGGGGTCTCTCTCCCTCTCTCTCTCTCTCT

CTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCCTCTCTCTCTCTCTCTTCTCTCTCTCTTCTCTCTCTCTCATCCCCTCGTTCCCTTCCTT

GCTTTCCTCCCCACTCCTCTC

HEMA154498.9

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 179 bp

Sequence 2: ref|AC_000066.1| 201 bp

Sequence 3: ref|AC_000155.1| 201 bp

ref|AC_000066.1| GGATTCCAGTGACATTGACTCTATCATTCTTTTACCCCTATTCTTCCCTGCACATTTATC

ref|AC_000155.1| GGATTCCAGTGACATTGACTCTATCATTCTTTTACCCCTATTCTTCCCTGCACATTTATC

ref|NC_000023.9| GGATTCCAGTGACATTGACTCTATCATTCTTTTACCCCTATTTTTCCCTGCACATTTATC

****************************************** *****************

ref|AC_000066.1| TGGGGTCTCTCTCCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCT

ref|AC_000155.1| TGGGGTCTCTCTCCCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCTCT

ref|NC_000023.9| TGGGGTCTCTCTC----------------------TCTCTCTCTCTCTCTCTCTCTCCTC

************* **********************

ref|AC_000066.1| CCTCTCTCTCTCTCTCTTCTCTCTCTCTTCTCTCTCTCTCATCCCCTCGTTCCCTTCCTT

ref|AC_000155.1| CCTCTCTCTCTCTCTCTTCTCTCTCTCTTCTCTCTCTCTCATCCCCTCGTTCCCTTCCTT

ref|NC_000023.9| TCTCTCTCTCTCTCTCTTCTCTCTCTCTTCTCTCTCTCTCATCCCCTCGTTCCCTTCCTT

***********************************************************

ref|AC_000066.1| GCTTTCCTCCCCACTCCTCTC

ref|AC_000155.1| GCTTTCCTCCCCACTCCTCTC

ref|NC_000023.9| GCTTTCCTCCCCACTCCTCTC

*********************

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
HEMA154507.3	NC_000023	154507351--154507428	20.5	87	119
	AC_000066	155007871--155007952	21.5	87	127
	AC_000155	143335156--143335241	22.5	88	135

New!	This microsatellite marker has been recently validated experimentally as informative^[1].
New!	Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD) ^[1]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect)^2].
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.
^[2]	Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.

HEMA154507.3	Information
Amplicon	356-364 bp
Forward ampliprimer	GGGATTCATCTGGCATTTATTT
Reverse ampliprimer	GGGATTTTGTAGTGTTGCTATGG

>ref|NC_000023.9|

GGGATTCATCTGGCATTTATTTACAATGAGTTTCCTTTTCAACTAACTACCATACACCAGGTAACCTGAGCATATCTACAAATCTTTTAA

CAAAATTGTTATACAAATAATGTATGTTTTAGATGAAAAACTAGAAATTACAGAAAAACACAAATGAAGAAAAGTTAAATATTTATACTT

TTACCACTTTCTTACAGAAATACATTATTTACCTATCTGTATCTATATAATCTATATTTGCATCTATCATCTATTTATCTATCATCTATC

TATCTATCTATCTATCTATCTATCTATCTATCTATCATCTATCATCTATACATCTATTTGTGGCCATAGCAACACTACAAAATCCC

>ref|AC_000066.1|

GGGATTCATCTGGCATTTATTTACAATGAGTTTCCTTTTCAACTAACTACCATACACCAGGTAACCTGAGCATATCTACAAATCTTTTAA

CAAAATTGTTATACAAATAATGTATGTTTTAGATGAAAAACTAGAAATTACAGAAAAACACAAATGAAGAAAAGTTAAATATTTATACTT

TTACCACTTTCTTACAGAAATACATTATTTACCTATCTGTATCTATATAATCTATATTTGCATCTATCATCTATTTATCTATCATCTATC

TATCTATCTATCTATCTATCTATCTATCTATCTATCTATCATCTATCATCTATACATCTATTTGTGGCCATAGCAACACTACAAAATCCC

>ref|AC_000155.1|

GGGATTCATCTGGCATTTATTTACAATGAGTTTCCTTTTCAACTAACTACCATACACCAGGTAACCTGAGCATATCTACAAATCTTTTAA

CAAAATTGTTATACAAATAATGTATGTTTTAGATGAAAAACTAGAAATTACAGAAAAACACAAATGAAGAAAAGTTAAATATTTATACTT

TTACCACTTTCTTACAGAAATACATTATTTACCTATCTGTATCTATATAATCTATATTTGCATCTATCATCTATTTATCTATCATCTATC

TATCTATCTATCTATCTATCTATCTATCTATCTATCTATCTATCATCTATCATCTATACATCTATTTGTGGCCATAGCAACACTACAAAA

TCCC

HEMA154507.3

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 356 bp

Sequence 2: ref|AC_000066.1| 360 bp

Sequence 3: ref|AC_000155.1| 364 bp

ref|NC_000023.9| GGGATTCATCTGGCATTTATTTACAATGAGTTTCCTTTTCAACTAACTACCATACACCAG

ref|AC_000155.1| GGGATTCATCTGGCATTTATTTACAATGAGTTTCCTTTTCAACTAACTACCATACACCAG

ref|AC_000066.1| GGGATTCATCTGGCATTTATTTACAATGAGTTTCCTTTTCAACTAACTACCATACACCAG

************************************************************

ref|NC_000023.9| GTAACCTGAGCATATCTACAAATCTTTTAACAAAATTGTTATACAAATAATGTATGTTTT

ref|AC_000155.1| GTAACCTGAGCATATCTACAAATCTTTTAACAAAATTGTTATACAAATAATGTATGTTTT

ref|AC_000066.1| GTAACCTGAGCATATCTACAAATCTTTTAACAAAATTGTTATACAAATAATGTATGTTTT

************************************************************

ref|NC_000023.9| AGATGAAAAACTAGAAATTACAGAAAAACACAAATGAAGAAAAGTTAAATATTTATACTT

ref|AC_000155.1| AGATGAAAAACTAGAAATTACAGAAAAACACAAATGAAGAAAAGTTAAATATTTATACTT

ref|AC_000066.1| AGATGAAAAACTAGAAATTACAGAAAAACACAAATGAAGAAAAGTTAAATATTTATACTT

************************************************************

ref|NC_000023.9| TTACCACTTTCTTACAGAAATACATTATTTACCTATCTGTATCTATATAATCTATATTTG

ref|AC_000155.1| TTACCACTTTCTTACAGAAATACATTATTTACCTATCTGTATCTATATAATCTATATTTG

ref|AC_000066.1| TTACCACTTTCTTACAGAAATACATTATTTACCTATCTGTATCTATATAATCTATATTTG

************************************************************

ref|NC_000023.9| CATCTATCATCTATTTATCTATCATCTATCTATCTATCTATCTATCTATCTATCTATCTA

ref|AC_000155.1| CATCTATCATCTATTTATCTATCATCTATCTATCTATCTATCTATCTATCTATCTATCTA

ref|AC_000066.1| CATCTATCATCTATTTATCTATCATCTATCTATCTATCTATCTATCTATCTATCTATCTA

************************************************************

ref|NC_000023.9| TCTATC--------ATCTATCATCTATACATCTATTTGTGGCCATAGCAACACTACAAAA

ref|AC_000155.1| TCTATCTATCTATCATCTATCATCTATACATCTATTTGTGGCCATAGCAACACTACAAAA

ref|AC_000066.1| TCTATCTA----TCATCTATCATCTATACATCTATTTGTGGCCATAGCAACACTACAAAA

****** **********************************************

ref|NC_000023.9| TCCC

ref|AC_000155.1| TCCC

ref|AC_000066.1| TCCC

****

*Locus* name	Reference X-chromosome	Localization	Copy number	(%)Match	Score
DXS1108	NC_000023	154515075--154515113	19.5	94	69
	AC_000066	155015601--155015639	19.5	94	69
	AC_000155	143342903--143342953	25.5	95	93

New!

This locus has recently been tried-and-tested as informative using the pair of primers here provided^[1].

^[1]

Reitter-Pfoertner S, von Haeseler A, Horvath B, Sunder-Plassmann R, Tiedje V, Pabinger I, et al. Identification of an ancient haemophilia A splice site mutation. Thromb Res 2012; 130: 445-450.

DXS1108	Information
Amplicon	118-130 bp
Forward ampliprimer	GGGAGATAGGAATGATGGAGTG
Reverse ampliprimer	TATTTTCTGGGCCATCTTGG

>ref|NC_000023.9|

GGGAGATAGGAATGATGGAGTGAATTCATCATATGTGATTTCCACAGCCACCACACACACACACACACACACACACACGCACACACACAC

TGTATCCCCCAAGATGGCCCAGAAAATA

>ref|AC_000066.1|

GGGAGATAGGAATGATGGAGTGAATTCATCATATGTGATTTCCACAGCCACCACACACACACACACACACACACACACGCACACACACAC

TGTATCCCCCAAGATGGCCCAGAAAATA

>ref|AC_000155.1|

GGGAGATAGGAATGATGGAGTGAATTCATCATATGTGATTTCCACAGCCACCACACACACACACACACACACACACACACACACACACAC

GCACACACACACTGTATCCCCCAAGATGGCCCAGAAAATA

DXS1108

In-Silico polymorphism validation

Sequence 1: ref|NC_000023.9| 118 bp

Sequence 2: ref|AC_000066.1| 118 bp

Sequence 3: ref|AC_000155.1| 130 bp

ref|NC_000023.9| GGGAGATAGGAATGATGGAGTGAATTCATCATATGTGATTTCCACAGCCACCACACACAC

ref|AC_000066.1| GGGAGATAGGAATGATGGAGTGAATTCATCATATGTGATTTCCACAGCCACCACACACAC

ref|AC_000155.1| GGGAGATAGGAATGATGGAGTGAATTCATCATATGTGATTTCCACAGCCACCACACACAC

************************************************************

ref|NC_000023.9| ACACACACACACACACAC------------GCACACACACACTGTATCCCCCAAGATGGC

ref|AC_000066.1| ACACACACACACACACAC------------GCACACACACACTGTATCCCCCAAGATGGC

ref|AC_000155.1| ACACACACACACACACACACACACACACACGCACACACACACTGTATCCCCCAAGATGGC

****************** ******************************

ref|NC_000023.9| CCAGAAAATA

ref|AC_000066.1| CCAGAAAATA

ref|AC_000155.1| CCAGAAAATA

**********

Table S3. Primers for amplification of complete set of STR loci.

Each selected STR locus, masked and flanked, was run in the online software *OligoPerfect™ Designer* from Invitrogen™, with parameters set for multiplex PCR assays: application (PCR detection), primer size (18-27 bases), primer Tm (57-63 ^oC), primer %GC (20-80), product size (100-450 bp), primer concentration (50 nM), salt concentration (50 mM). Absence of structure constraints (potential hairpin formation, 3´ complementary, potential self-annealing sites) was confirmed using the software *Oligo Calc: Oligonucleotide Properties Calculator. We validated In-Silico* the uniqueness of each amplicon sequence using Blat (UCSC), which quickly maps sequences to the genome reference assemblies, i-PCR (UCSC), Primer-BLAST (NCBI), WebSat, and Multiple Sequence Alignment by CLUSTALW.
n^o	*Locus* name	Forward sequence (5’-3’)	Reverse sequence (5’-3’)	Size range (bp)
1	DXS9897	TTCTGCTGTGCAATACATCTGA	CAGCAGACATTATTGAGGGAGA	232-236
2	HEMA152128.2	GCACATACAGCCTTCAAGAGG	CGATGAAAAGTCACTGCGTCT	220-222
3	HEMA152141.6	GGCAGCCACAGGTAAGACAT	CTTGAACTTGGGAGGTGGAG	259-261
4	HEMA152149.9	TGGGCTTTATTCCCTTCCAT	TGACGAGTTAGTGGGTGCAG	260
5	HEMA152155.2	TGTTGTCATCCTGTGAGTCAA	GCAAAGGATTCTTCCACCAG	260-262
6	HEMA152155.6	TGGATGTACATTTCCCATTTCC	TCTGTTTGCCTGTGAAGCAC	193-197
7	LOC649201Int2	TTGCCTTGATTGTGGTGATG	GTTCACGTCTTTTGCCCATT	215-231
8	HEMA152257.3	TTTCAACAGTTAATCATGCCAAT	TGCTGCCTTTAAACCATCCT	246-260
9	DXS15	CATACGCAATAAATAGCACATGGT	AGAGAATGAAGAAATTAGGTGCTGA	250
10	HEMA152317.9	TCCTGTCCTTCATTCCTGAT	AACCACGTGTGTTAGCATGTG	251
11	HEMA152342.0	GGAGAATGCTGCTTGGAGTG	AGGATGGGGAATAGGGACAC	183-185
12	DXS1356	TTCAGTGAGCCAAGATCACG	GAGGGGTCTTCTGGCTCTTC	286
13	UCHL5IPInt8	TTGCTGCTTCTTCACGGTCT	GGGTGGGAAGCAGGAGAT	168
14	DXS9901(BGNExon8)	CTTCGACCAGTCCTCCCTTC	AGAAAGCAGTCCCAGCTCAA	250-252
15	HEMA152442.8	GGCTTAGAAAATCCCTTTCG	GAAAAAGCTTGCTAATCCCTAAA	164
16	ATP2B3Int20	CCCTCCTCTTGCATAAGCTG	ACTGCTTTTCAGCCCAACTG	251-257
17	HEMA152503.2	CTCCAGCCTGAGTGACAGAG	ACACCAATGCAGCAAGGAC	234-254
18	HEMA152520.9	ATCCACAGCCTCCATCTCTG	GGGAGGTGGAGGTTACAGTG	179
19	HEMA152524.1	AAAATCCCAGCTGGCTTAAA	TCAGCTTGTCAATGTCTGCAC	257-292
20	HEMA152539.8	GGTTGTGGTGAGCCAAGATT	TGGGTGACAGAGCAAGACTG	254
21	DXS8087(SHGC-152185)	CTGCGCCAGTGAACAAGG	GGGTCATGCACAAATGCAC	130-132
22	HEMA152581.8 *WARNING!*	GGATTACTTGAGCCCAGGAG	CCACCATGCTCGGTTCTAAT	132
23	PNCKInt2	CATCATCTCACACCCACTCG	AGACATGCTGCTGCTGAAGA	179
24	SLC6A8Exon1	GACGGGGAAGAGAGGGATAG	AGCCTGAAGAATCTCATGTGTG	439
25	PDZD4Int1	CCAACTCCTGTGCAAAATGA	GAGTGGGAGGATTGCTTGAA	235-237
26	REN87871^(j)*WARNING!*	TCGCCTCGCTAATTTTTCTT	CTGGGCAACATAGCAAGACC	101-120
27	HEMA152778.1	CTCTAGCCTGGGTGATCGAG	TCAGCTGAAGACCCATTTCC	232
28	REN87914	AAGTGGAGGGCTCACCTGT	GCGAGACAAAGAAAGAAAAAGAA	350-360
29	DXS9796(REN88038)	CGCAGCATACTTTTTGAGCTT	ACAAACCCTGATCCAACCAA	153
30	REN88052	TGCTGCTTCCTCCTTGTTTT	GCTGTTGAGTGTCCTCACGA	100-102
31	REN88058	TCCAACCTGGGTGACAGAGT	TTCTGGTGAGGGCTGTCTTC	279-287
32	RENBPInt9.1	CTGCACTGAGCCATGATTGT	CCTGTGTCATGTCAAGCAGTC	193
33	REN88388(TMEM187Int1.1)	GAGCAAGACCCTGTCTCAAAA	ACCTCCAGGCCTTTAGCATT	200
34	TMEM187Int1.2	GAGTGCAGTGGCGTGATCT	GCGGCAGAGTGAGACTTCAT	190-195
35	REN88442	CTGCCTCAGCCTCCAGAGTA	GGTTGCAGTGAGCCAATAGC	185
36	HEMA152913.2	TGAGAAGACGTGGACACTGC	CCTGGGAGAAAGGGTGAGAC	254-274
37	REN88627(MECP2Int2.3)	GGCGAACAAACCAGACACAT	ATGCAGTAAGCCGAGATTGC	205-225
38	REN88666/HSC270_(M3)/(MECP2Int2.2)	GAATAGGCATCTGAGGACAGC	TGTGCTTTGTGACATGATGG	148-150
39	REN88881	CACAGACACAGACACGCAGA	TGTGTATGATTGGAATTGCCTAC	141-143
40	HEMA153020.0	GGAGGTGGAGGTTGCAGTAA	TTTGCCCTCTCCTAACCTTTT	193-205
41	REN88942	TTGCTCCATTAGCTCCAGGT	GGGCCCTTCCTTCTCTCTAA	235
42	REN88956	CCTCAGCTCCCCAAGTAGC	GCAGGCAGATTGCTTGAACT	172
43	HEMA153047.7	GCTCACAAGTGTAATCCCAGTG	TCAAATGATCCTCCCACCTC	272-280
44	HEMA153050.0	GTGTTGCCCAGCCTACTCTC	CCTGAGAGACAGAGGTTGCAG	286
45	TKTL1Int1	ACACCCTGACTCGTTTCCTG	CCAGGAGGTCAAGGCTGTAG	199-215
46	REN89483(TAZInt5)	AGGGCTGGTCTTGAACTCCT	CACCCCACCTGGCTATCTAA	319-320
47	HEMA153333.1	CACCTGGCGTCACTAGGAAT	CTGGGAGACAGAGCAAGACC	182
48	G65800	TCCAAAGTGCTGGGATTACA	AGGGTGCAGTGAGCTGAGAT	168
49	REN89870(FAM3AInt2)	TCATGCCTGTAGCACTTTGG	CCTGGGTTCAAGCGATTCTA	233-236
50	REN89898/REN89899	GATTGCTTGAAGCCAGGAAC	TGTGGAGTCTAGCTGCCCTTT	120-122
51	stSG604115/REN89922	GGCAACTTGGGGGTTAATTT	AGGAGACTGAGGTGGGAGGT	211
52	REN90010/GDB:179459/(IKBKGInt1) New!	ATCGGTTGGGGGATACATTT	TCTACCCAAGGCCTTCTTTG	183
53	stSG604170/REN90051/REN90052	ATGGGCTTGGACTTGGTATG	AGCAGCTCCTCACCAGGAT	318
54	DXS1073(REN90074) *WARNING!*	AAGAATGCCCTCTCCGAGTT	ATTGGTGGCCTTTGAAACAC	132-134
55	REN90085	ACCATTCCCTCCCTGACTCT	GCCAGACCCCATCTCTCAAG	223
56	REN90200(HA472)	GCTCCTTTGATTGGATAATTTCA	TGCCTCAACATCAGAATAGACC	256-269
57	stSG604240/REN90371 (GAB3Int2.2)	CTTCTTTATGGCCCTCAGCA	TCTGGGCACATGGTAGATGTAA	219-227
58	stSG604242/REN90378 (GAB3Int2.1)	AATGTCAGGGGCTCAACAAC	TAAGTTTGCTCTGGGCCATT	215
59	stSG604248/REN90414 (GAB3Int1) New!	CCTGGGCAACAGAGTGAGAC	GAAGGGGAGAAGCAGTTTCC	147
60	stSG604294/REN90682/ UniSTS:68640/HSC269_(M1)(MPP1Int5)	GGATCTGGGTCGTCTGTCTC	GAGTTGGGAAGTGCCATCAT	273-281
61a	REN90833 New!	GCAAATTGTGAGGGAGGTATG	GAACTGCATTGGGCAAATCT	118-130
61b	REN90833 (redesigned)* *WARNING!*	GGCAAATTGTGAGGGAGGTA	CCAAAGGGAAAAGTCAAGCA	138-160*
62	REN90932(F8Int25.3)	TCCAAGATCAAGGGGTAGGC	GCCTGGACTACAGAGGGAGA	122
63	stSG604314/REN90841(F8Int25.2) New!	CTCTAGGCTGCCAACTCACC	TCATGGTTCCCAAAGAAACA	144-148
64	REN90957(F8Int25.1)	CTGCCTTTGGACTGGAACAT	TGTGTTAGTCAGGGATATCCAAA	145
65	REN90965(F8Int24.1)	ATGAGGTTTCACCAGGTTGG	GGCAGGATAATCGCATGAAC	287
66	F8Int22 New!	AAGACCCTTAGCTGTTTCATAAGC	TTCATACAGTGGGATCATTCATT	221-223
67	HSC271_(M4)/F8Int21 New!	AGCCTGCTTCTTGACAAACC	AGGTGTGGGCAAAGCATTAG	164
68	REN91100(F8Int13.2) New!	GCCTAGAGAATGCCAAAGTAACA	TTGTTGTAATTCCCATTTGCAT	148-150
69	REN91114(F8Int13.1)	GCCAACACACACTGAAGGAA	CTCCCAATGCAAAGACCTTC	264-266
70	stSG604366/REN91179/REN91180(F8Int10)	TGAATATGCGAAGAACCCTGA	CGGTTTTCAGCTACATTCCAC	201
71	stSG604368(F8Int9.2) New!	CTGGGTGACAGAATGAGACC	TCCTCATATCACACACCTCAGA	165-166
72	stSG604369/REN91192 (F8Int9.1)	AGATTCGAGCGATTCTCCTG	CAGTCATTGCTGTGGGTTTG	233
73	REN91235(F8Int6)	TTCTCCTGCTTCAGCCTCTC	AGCATATCCACCCTCACCAC	325
74	stSG604384/REN91318 (F8Int3)	AGCAGGAAGAAATAAAATTGTCC	CTGAGGCAGGAAAATCACTTG	221
75	F8Int1	CTGCCCTTGGACATAAGCAT	CCATATGATCCAGCAACTCG	218-220
76	HEMA154006.7	CATGGAAAGGAGGAAGGGTA	TGGACGGAGCTGAGACAGAT	274-276
77	stSG604469	TCATCGTGCTAATGTGCATGT	TATGTGGCCTGCTGAGAATG	114-136
78	stSG604486	TGGCAATTTCAGATCACTTCTTT	GGTTTTCTTTGTGGCCAAGT	204-216
79	VBP1Int4	GGTCAGTTTCCCCCAGGTAT	CAAAGCAGGGGACCAAAAA	183
80	HEMA154126.0	TTTCACTGCCTTGGTTAAATG	CAGAGGGTACATGTGCAGGTT	274-275
81	HEMA154130.5	ATGATGTACAAGCCCTACCTGA	TGAGCCAGGGAAGAATGAAA	162
82	CLIC2Int1	GCACATGCATCCAAGAACTG	CTCCAATTCCTTCCAGGTTG	173-177
83	HEMA154229.0 *WARNING!*	AAGCCTGGGAGGATGGACTA	GGGCGACAGAGCGAGACT	96-100
84	HEMA154279.5(HA544)	AAAGTGAAACTGTCCGTGTTTG	TTGCAGTGAGGTGAGATTGC	157-175
85	HEMA154281.2	TCTGCACTCCCATGTTCATT	CTGTCCACTCCCCTACTCCA	378
86	HEMA154311.3 New!	GTTTTGGTCCTGGCACTCAT	TTCACGGTTTTAATTTTTCATGTC	254-269
87a	TMLHEInt5 New!	CCAAAACTAATGGTTAGGCAAT	GAGGGATAGCATCGGGAGAT	276-286
87b	TMLHEInt5 (redesigned) **WARNING! ****	TCCAAAACTAATGGTTAGGCAAT	CACCAGCATGGCACATGTAT	219-244**
88	TMLHEInt2	TGTCCTAGGGTGTGTGGAAT	CACTCTAGGGAAAGGGTTCTCA	133-136
89	TMLHEInt1.3 New!	ATCTTCCTCCCTCAAGTGCT	ATGATCATGCCACTGCACTC	165-169
90	TMLHEInt1.2	CAAAGGGGGATAATGAAGACA	TCTGTGTGCTCCAGGATTGA	296-300
91	TMLHEInt1.1 New!	GTAGTCCCAGGCACTCAGGA	CTGGGTTCTTTCCATTTCCA	247-251
92	HEMA154498.9	GGATTCCAGTGACATTGACTCTA	GAGAGGAGTGGGGAGGAAAG	179-201
93	HEMA154507.3 New!	GGGATTCATCTGGCATTTATTT	GGGATTTTGTAGTGTTGCTATGG	356-364
94	DXS1108	GGGAGATAGGAATGATGGAGTG	TATTTTCTGGGCCATCTTGG	118-130

*	REN90833 (redesigned). This microsatellite marker has been recently validated experimentally as informative,^[1] albeit using a different (redesigned) pair of primers, shown above. WARNING!* The redesigned REN90833 primers also anneal (reverse primer with 3bp 5’ mismatched to a non-STR locus on chromosome 10, yielding a 106 bp non-polymorphic PCR product with DNA from both males and females). This product does not interfere with designation of REN90833 alleles, which ranged 138-160 bp in the tested population.
**	TMLHEInt5 (redesigned). This microsatellite marker has been recently validated experimentally as informative,^[1] albeit using a different (redesigned) pair of primers. WARNING!** The TMLHEInt5 redesigned reverse primer matches a second location 1918 bp downwards, yielding a 2148 bp TMLHEInt5 specific extended product, and therefore polymorphic due to the STR element, which is excluded from analysis by capillary electrophoresis within the analysis range of 450 bp, and thus it does not interfere with TMLHEInt5 allele calling ranging 219-244 bp.
^[1]	Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.

Copyright and credits

Copyright © 2008

Original website coding and design by Enrique Medina–Acosta and Filipe Brum Machado.

Molecular Identification and Diagnosis Unit, Centre for Biosciences and Biotechnology State University of North Fluminense, Brazil.

HEMApSTR is maintained by:

Enrique Medina-Acosta, MSc, PhD.

Senior Associate Professor

Laboratory of Biotechnology

Universidade Estadual do Norte Fluminense Darcy Ribeiro - UENF

Head of the Molecular Identification and Diagnosis Unit (Núcleo de Diagnóstico e Investigação Molecular – NUDIM)

Avenida Alberto Lamego 2000, Parque Califórnia, Campos dos Goytacazes, RJ, CEP 28013602, Brazil.

Email: quique@uenf.br

Suggested citation:

FORENSIC APPLICATIONS OF ChrX-STRs:

With the overwhelming availability of human genome sequence reference public databases and the rapid development of bioinformatics tools to ease In-Silico identification, characterization and comparison of DNA sequence features, the number of polymorphic sequences, namely, single nucleotide polymorphisms (SNPs), short tandem repeats (STRs) and copy number variations (CNVs) relevant for medical and forensic genetics has steeply increased within the last few years ^[1]. Thanks to massive interest on human disease association studies, comprehensive physical and genetic maps for about 3.1 million of genotyped SNPs are public-available through the International HapMap Project ^[2]. Genome wide information on gametic linkage disequilibrium (GD) between linked and unlinked markers can be easily retrieved from different genome browsers (i.e., Ensembl; dbSNP; James Watson Genome Browser; FastSNP; HapMap; Bushman (Complete Khoisan and Bantu genomes from southern Africa). Unfortunately, similar level of information regarding STRs is lagging considerably and where existent it is not readily available as a comprehensive public database or in easily retrievable flat dumping formats. This is clearly the case for the STRs markers currently used in forensic practice, whether located on autosomes or sex chromosomes. Concerning STRs on human X-chromosome (X-STRs) there are prospects of comprehensive combined physical and genetic maps through validation In-Silico ^[3]. An initiative to compile reported uses in forensics and evolutionary anthropology is available at ChrX-STR (for those interested in databases that cover strictly forensic issues concerning the usage of X-chromosomal markers), but currently does not offer a comprehensive and updated dataset downloadable in any format. Moreover, there are conflicting results regarding the genetic map distances between typing X-STRs and the occurrence of truly stable segregating haplotype blocks ^[4-10]. Hence, it is unclear to what extent that inferable “stable” haplotype block assessment precludes forensic usefulness of STRs assays for the resolution of so called sample-deficient complex kinship cases, as claimed ^[4,11]. There is, therefore, a need for a reappraisal of the experimental evidence regarding genetic distances, recombination, and the occurrence of truly stable segregating haplotype blocks.

Attention has been called to the risks and consequences of the erroneous habit of applying the product rule to syntenic X-linked marker systems in the field of Forensic Science ^[15,16] (Visit Chromosome X-STRs for a thorough discussion on combined power of discrimination in X-linked STRs). Meaningful calculation of Combined Power of Discrimination requires accurate knowledge about genetic linkage status of typing loci. The inappropriateness of the product rule for syntenic loci is not a new issue. However, aware of the problem, many forensic researches working with linked X-chromosomal markers fail to curtail misuse of the product rule. By ignoring linkage disequilibrium, typing laboratories erroneously claim that their X-STRs marker sets offer comparable, if not greater, discriminatory power than typing with an equal number of unlinked autosomal markers that exhibit similar power of discrimination values. Considering the genetic map length (196 cM) of the human X chromosome, the product rule is suitable only to compute combined probabilities from no more than four points (whether single loci or groups of contiguous loci, the latter in extensive gametic disequilibrium), each point mapping >50 cM apart, which underlines independent segregation. Thus, for the markers commonly used in forensic practice, it urges to identify specific multiallelic interallelic associations, their frequency and strength, which can be measured, for example, using the D' coefficient or r² statistics. We caution that tests of statistical significance (P values) are not measures of gametic disequilibrium.

No direct and comprehensive experimental (i.e., DNA typing in extended pedigrees) evidence currently exits to accurately determining recombination rates between any pair of forensic X-STRs markers. Thus, all studies refer to genetic distances determined for neighboring (or contiguous) SNPs markers in linkage and/or association studies in various populations ^[12-14] to infer, and therefore assume at risk of considerable linkage error, whether there is or not evidence of recombination.

For a comprehensive definition of gametic disequilibrium, readers are suggested to read Medina-Acosta, E. (2013) Gametic disequilibrium. In: Maloy S and Hughes K (ed.) Brenner's Encyclopedia of Genetics, vol. 3, pp. 151-153. Elsevier: New York (authors copy available at Chromosome X-STRs).

References (for Forensic Applications of ChrX-STRs):

1.	Oetting WS. Clinical genetics & human genome variation: the 2008 Human Genome Variation Society scientific meeting. Hum Mutat 2009; 30(5): 852-6.
2.	Frazer KA, Ballinger DG, Cox DR, Hinds DA, Stuve LL, Gibbs RA et al. A second generation human haplotype map of over 3.1 million SNPs. Nature 2007; 449(7164): 851-61.
3.	Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.
4.	Szibor R, Hering S, Kuhlisch E, Plate I, Demberger S, Krawczak M et al. Haplotyping of STR cluster DXS6801-DXS6809-DXS6789 on Xq21 provides a powerful tool for kinship testing. Int J Legal Med 2005; 119(6): 363-9.
5.	Hering S, Augustin C, Edelmann J, Heidel M, Dressler J, Rodig H et al. DXS10079, DXS10074 and DXS10075 are STRs located within a 280-kb region of Xq12 and provide stable haplotypes useful for complex kinship cases. Int J Legal Med 2006; 120(6): 337-45.
6.	Becker D, Rodig H, Augustin C, Edelmann J, Gotz F, Hering S et al. Population genetic evaluation of eight X-chromosomal short tandem repeat loci using Mentype Argus X-8 PCR amplification kit. Forensic Sci Int Genet 2008; 2(1): 69-74.
7.	Edelmann J, Hering S, Augustin C, Szibor R. Characterisation of the STR markers DXS10146, DXS10134 and DXS10147 located within a 79.1 kb region at Xq28. Forensic Sci Int Genet 2008; 2(1): 41-6.
8.	Hundertmark T, Hering S, Edelmann J, Augustin C, Plate I, Szibor R. The STR cluster DXS10148-DXS8378-DXS10135 provides a powerful tool for X-chromosomal haplotyping at Xp22. Int J Legal Med 2008; 122(6): 489-92.
9.	Tillmar AO, Mostad P, Egeland T, Lindblom B, Holmlund G, Montelius K. Analysis of linkage and linkage disequilibrium for eight X-STR markers. Forensic Sci Int Genet 2008; 3(1): 37-41.
10.	Machado FB, Medina-Acosta E. Genetic map of human X-linked microsatellites used in forensic practice. Forensic Sci Int Genet 2009; 3(3): 202-4.
11.	Edelmann J, Hering S, Augustin C, Kalis S, Szibor R. Validation of six closely linked STRs located in the chromosome X centromere region. Int J Legal Med 2009.
12.	Nagaraja R, MacMillan S, Kere J, Jones C, Griffin S, Schmatz M et al. X-chromosome map at 75-kb STS resolution, revealing extremes of recombination and GC content. Genome Res 1997; 7(3): 210-22.
13.	Stassen HH, Scharfetter C. Integration of genetic maps by polynomial transformations. Am J Med Genet 2000; 96(1): 108-13.
14.	Matise TC, Chen F, Chen W, De La Vega FM, Hansen M, He C et al. A second-generation combined linkage physical map of the human genome. Genome Res 2007; 17(12): 1783-86.
15.	Medina-Acosta, E., Evidence of partial and weak gametic disequilibrium across clusters of pericentromeric short tandem repeats loci on human X chromosome: Proceed with caution in forensic genetics. Forensic Sci Int Genet, 2010. DOI 10.1016/j.fsigen.2009.12.002 (published online ahead of print).
16.	Medina Acosta, E. Machado, BF. Eyes wide open: The (mis)use of combined power of discrimination for X-linked short tandem repeats. Mol Biol Rep 2001; 38(1):4003-4006.

Version updates (changes, clarifications, and WARNINGS!)

HEMApSTR version 1.0: April 4, 2008 (includes gene links to NCBI).

HEMApSTR version 1.1: July 27, 2008 (includes gene links to OMIM).

HEMApSTR version 1.2: August 12, 2008 (includes links to LIFEdb Xq28).

HEMApSTR version 1.2: September 12, 2008. The following WARNING was included: The published primer pair sequences ^[1] anneal to more than one location on contig(s) by e-PCR: chrX:154228869-154229259 and chrX:154377350-154377732 on human chromosome X. The locations are identical at each side of the STR element in over 150 nucleotides. Therefore, the marker HEMA154229.0 cannot be considered unique (UniSTS).

^[1]Machado FB, Medina-Acosta E. High-resolution combined linkage physical map of short tandem repeat loci on human chromosome band Xq28 for indirect haemophilia A carrier detection. Haemophilia 2009, 15:297-308.

HEMApSTR version 1.2: February 2, 2009. HEMApSTR site begins to be accessed from NCBI F8 gene link.

HEMApSTR version 1.3: May 22, 2009 (New microsatellite markers validated experimentally as informative by heterozygosity rates).

Machado FB, Duarte LP, Medina-Acosta E. Improved criterion-referenced assessment in indirect tracking of haemophilia A using a 0.23 cM-resolution dense polymorphic marker set. Haemophilia 2009, 15:1135-42.

HEMApSTR version 1.4: September 30,2009. The following WARNING was included: some pairs of primers may form dimers and, therefore, their inclusion in multiplex assays may cause unbalanced profiles. This effect, however, has not been observed with the published marker sets. Example of dimmer formation (Analyzed using online Autodimer program): Primer Dimer Results for the heptaplex PCR assay reported by Machado et al. (Haemophilia 2009; 15(5):1135-42).

Sequences	Match Sequence	Matches	Score	Est. Tm (°C)	ΔG(@ 37.0 °C) (kcal/mol)	Lineup
HEMA154507.3R GGGATTTTGTAGTGTTGCTATGG vs. F8Int25.2R TCATGGTTCCCAAAGAAACA	CCANAGNAACA	9	7	< 0	-2.5	3'-GGTATCGTTGTGATGTTTTAGGG-5' \|\|\|x\|\|x\|\|\|\| 5'-TCATGGTTCCCAAAGAAACA-3'
HEMA154507.3R GGGATTTTGTAGTGTTGCTATGG vs. F8Int13.2F GCCTAGAGAATGCCAAAGTAACA	TGTTNCTNTGG	9	7	< 0	-2.5	3'-ACAATGAAACCGTAAGAGATCCG-5' \|\|\|\|x\|\|x\|\|\| 5'-GGGATTTTGTAGTGTTGCTATGG-3'
F8Int13.2F GCCTAGAGAATGCCAAAGTAACA vs. F8Int25.2 CTCTAGGCTGCCAACTCACC	GCCTAGAG	8	8	17.8	-5.6	5'-GCCTAGAGAATGCCAAAGTAACA-3' \|\|\|\|\|\|\|\| 3'-CCACTCAACCGTCGGATCTC-5'

HEMApSTR version 1.5: October 25, 2009. A WARNING NOTE ON FORENSIC APPLICATIONS OF ChrX-STRs was included.

HEMApSTR version 1.6: March 6, 2010. The following information and WARNINGS were included:

Extended haplotype homozygosity analysis centered at the F8 intronic STR loci has revealed that the most frequent core haplotypes decay the least in the flanking gametic disequilibrium (GD)^[1]. The F8 intronic loci attend distinct non-random association forces; F8Int13.2 serves at maintenance of the long-range overlapping pattern of GD, whereas F8Int25.2 and F8Int22 serve at lessening it in force or effect)

^[1] Medina-Acosta E. Interlocus non-random association of multiallelic polymorphisms spanning the coagulation factor VIII gene on human chromosome distalmost Xq28. Haemophilia 2010, 16:525–537.

The published forward primer sequence (GGCAGGAGGATTACTTGAC) is missing a G residue at the 3’ end, and it is therefore incorrect. The correct sequence is GGCAGGAGGATTACTTGAGC. this primer pair matches two locations on contig(s) from NCBI36/hg18 reference genome sequence by e-PCR: chrX:152581821+152581980 and chr15:24833223-24834846, but only one location on GRChH37/hg19 reference sequence: chrX:152928627+152928786.

The locus HEMA152581.8 is, however, unique (UniSTS) and it can be developed into a marker using the, otherwise, amended primer pair now listed.

To improve specificity (i.e., reducing the chances of amplification of head-to-head primers annealing to more than one location on contig(s) from NCBI36/hg18 reference genome sequence by e-PCR, the published^[1] primer pair sequences (CTCAACTCACAGCAGCCTCA and CTCTTGAGCCCAGGAGTTTG) are now amended to the primer pair listed in Table S3. The locus REN87871 can thus be developed into a UniSTS marker.

HEMApSTR site authors´ articles begin to be cited as GeneRIFs (Gene References Info Function at NCBI).

At least six other individual genome sequences (haploid and diploid) are currently available from public databases (Asian, Southern African Bushman (Complete Khoisan and Bantu genomes from southern Africa^[2]) and James Watson’s Genome); search date: 02/15/2010) and promising prospects of a deep catalogue of human genetic variation comprising 1000 individual genomes; we will make efforts to update the polymorphism indexes, as relevant STR information becomes available). For instance, in Table S2, validation indexes that include information about allelic variants sequenced in the Southern African Bushman^[2] are highlighted in green (i.e., 3); Validation indexes that include new allelic variants sequenced in the Southern African Bushman are highlighted in cyan (i.e., 1).

^[2]Schuster SC, Miller W, Ratan A, Tomsho LP, Giardine B, Kasson LR et al. Complete Khoisan and Bantu genomes from southern Africa. Nature; 463(7283): 943-7.

HEMApSTR version 1.7: August 12, 2010. The following information was included:

Multiallelic interlocus non-random association analysis revealed that GAB3Int1 is at gametic (linkage) EQUILIBRIUM with F8Int21, F8Int9.2, TMLHEInt1.3 or TMLHEInt1.1. The observed breakdown of gametic disequilibrium attests historical recombination between GAB3Int1 and the F8 gene.

Machado FB, Alves Da Silva AF, Rossetti LC, De Brasi CD, Medina-Acosta E. Informativeness of a novel multiallelic marker-set comprising an F8 intron 21 and three tightly linked loci for haemophilia A carriership analysis. Haemophilia 2011, 17:257-66.

HEMApSTR version 1.8: November 16, 2010. The following information was included:

Meaningful calculation of Combined Power of Discrimination (CPD)requires accurate knowledge about genetic linkage status of typing loci.

Medina Acosta, E. Machado, BF. Eyes wide open: The (mis)use of combined power of discrimination for X-linked short tandem repeats. Mol Biol Rep 2001; 38(1):4003-4006.

HEMApSTR version 1.9: June 18, 2011. The following information was included:

Visit Chromosome X-STRs for a thorough discussion on combined power of discrimination in X-linked STRs.

HEMApSTR version 2.0: September 15, 2012. The following information was included:

We warrant about the need to acknowledge accurate tracking genetic markers in haemophilia linkage analysis to avoid unnecessary priority claims.

Machado FB, Medina-Acosta E (2012) Accurate tracking genetic markers in haemophilia linkage analysis: a matter beyond scientific priority. Haemophilia 18:e359--e360

HEMApSTR version 2.1: September 15, 2013. The following information was included:

Reported on the use of SNPSTR combined markers, F8 intragenic, for haplotyping; reported that some F8 intragenic STRs are nor polymorphic, as predicted In-Silico.

de Sá Machado Araújo G, Louvain de Souza T, Medina-Acosta E. Use of SNPSTR combined markers for gametic phase determination of X chromosomes in Hemophilia A female carriers. In 58o. Congresso Brasileiro de Genética. 2012. Foz do Iguaçu: Sociedade Brasileira de Genética.

HEMApSTR version 2.2: April 4, 2017. The following information was included:

A tetradecaplex (13 STR loci plus the AMELX/AMELY indel locus) QF-PCR assay has been developed by Zhao e t al (2017), based on the STR information provided by HEMApSTR. The authors genotyped two population subsets (Chinese ad Caucasian) to report heterozygosity rates ranging from 0.43 to 0.84, and about 80% of individuals being heterozygous for at least five STR markers. The tetradecaplex assay was tried-and-tested valuable in the Preimplantation Genetic Diagnosis (PGD) of HEMA and in the in vitro fertilization (IVF)-PDG in an HEMA at risk couple.

Zhao M, Chen M, Tan AS, Cheah FS, Mathew J, Wong PC, et al. Single-tube tetradecaplex panel of highly polymorphic microsatellite markers <1 Mb from F8 for simplified preimplantation genetic diagnosis of Hemophilia A (2017) J Thromb Haemost (published ahead of time) (March 27) doi: 10.1111/jth.13685.

EMA-2008®