Probe set and kit for detecting dynamic mutation STR and VNTR genetic loci, and method

Abstract

Provided are a probe set and kit for detecting dynamic mutation STR and VNTR genetic loci, and a method. A detection range covers 73 genetic loci, involving 79 diseases. Long-fragment targeted capture and long-read sequencing technologies are used to detect target repeat region sequence information of all targeted genetic loci from a single sample at the same time, thereby overcoming the defects of conventional detection techniques including one detection for one disease, lack of target repeat region sequence information, a limited detection range, and a long time required for differential diagnosis.

Description

Probe set, kit and method for detecting dynamic mutation STR and VNTR gene sites Technical Field

[0001] The present invention relates to a detection probe group, detection equipment and kit for dynamically mutated STR and VNTR gene sites. Background Art

[0002] Currently, there are over 70 known dynamic mutation diseases, primarily involving the nervous system, with a total prevalence of approximately 1 in 1,000. While the spectrum of these diseases continues to expand with advances in molecular genetics, over 70% of these diseases remain undiagnosed. Given the diverse and widespread prevalence of these dynamic mutation diseases, the large number of genes implicated, the ongoing discovery of new genes, the diverse size and sequence conformation of pathogenic repeat expansions, and significant gaps in our understanding of their underlying biological mechanisms, there is a growing need for improved molecular detection methods for abnormal repeat expansions.

[0003] Currently, common diagnostic methods for detecting dynamic mutation diseases include Southern blot, repeat-primed PCR, and next-generation sequencing (NGS). Southern blot is complex, time-consuming, and labor-intensive, and lacks sequence information for the repeat region of the causative gene. Repeat-primed PCR, while simple and capable of indicating the presence of abnormal repeat expansions in dynamic mutation diseases characterized by over 100 or even 1,000 repeat expansions, cannot provide precise length and sequence information, and requires separate analysis using specific primers / probes for each gene.

[0004] Obtaining sequence information for the repeat region of a pathogenic gene facilitates disease analysis. For example, spinocerebellar ataxia type 1 (SCA1) is an autosomal dominant genetic disease caused by an abnormal CAG repeat expansion in the ATXN1 gene. The presence of a CAT interruption is important when diagnosing SCA1 patients with 39-44 repeat alleles, as only the uninterrupted allele is considered abnormal. Therefore, molecular diagnosis of such patients requires simultaneous analysis of the number of repeats of both CAG and CAT units. Second-generation NGS sequencing has some practicality in analyzing repeat expansions. Using STR bioinformatics algorithms, it can indicate whether the number of repeats in some genes exceeds the normal range, but a second validation approach is still required. Many large pathogenic repeat expansions, low-complexity sequences, and high-GC content fragments are difficult to analyze using NGS platforms, making second-generation NGS sequencing ineffective in exploring dynamic mutation-related variants.

[0005] The advent of third-generation long-read sequencing technology and advances in computational analysis have significantly accelerated the discovery of dynamically mutated disease-associated genes. Between 2019 and 2021 alone, 17 new pathogenic and risk-associated tandem repeat expansions were reported. Third-generation sequencing provides a powerful tool for discovering longer and more complex repeat sequences.

[0006] In 2023, Hope Group released its third-generation dynamic mutation detection chip, "Repeat Hunter," and published the related patent CN116042610A. According to the patent, first, in terms of probe design, the probe design extends 3kb upstream and downstream of the STR locus, with an average spacing of 1000bp between each two adjacent probes. This probe design approach may not be suitable for regions with high homology or abnormal GC content, thereby affecting the capture efficiency of individual genes. Second, regarding genomic DNA fragmentation, the fragmented DNA fragments are 4.5-5.5kb. Genomic DNA is randomly fragmented and requires flanking regions for gene mapping. The shorter the fragmented DNA, the less template is available for the target repeat region, including the flanking regions. Third, regarding sequencing library length, a qualified library requires a main band length of at least 2kb. Hope Group performs two PCR amplifications during the library construction process. Because shorter fragments have greater amplification advantages, these two PCR amplifications may result in a main band of less than 5kb. Currently, literature has reported that the abnormally amplified repeat region of the PLIN4 gene can reach more than 4kb. Therefore, increasing the length of the main band of the library is conducive to detecting samples with extremely large abnormal repeat amplification.

[0007] Summary of the Invention

[0008] Given that the current third-generation long-read sequencing technology can meet the urgent need for rapid, accurate and comprehensive molecular diagnosis of repetitive amplification of dynamic mutation diseases, and overcome the shortcomings of traditional detection technology such as single disease single detection, no target repetitive region sequence information, small detectable range and long time required for differential diagnosis, the present invention provides a technology that can simultaneously detect multiple dynamic mutation diseases and provide target repetitive region sequence information, in order to assist clinicians in making differential diagnoses faster and more accurately and shorten the time to diagnosis.

[0009] The inventors discovered that the human genome contains over one million annotated tandem repeat sequences. Due to their repetitiveness and abundance in the human genome, target repeat regions may not be suitable for probe design. Furthermore, abnormal repeat expansion samples often contain high GC or high AT regions, making detection difficult. Therefore, probes were designed to flank the STR and VNTR loci, using specific sequences from both flanking regions to capture the target repeat regions.

[0010] The inventors have discovered that within the spectrum of dynamic mutational diseases, pathogenic repeat units are diverse, and the number of repetitions of the same unit varies significantly between individuals, sometimes as high as a thousand times. Therefore, longer random fragments of genomic DNA, longer PCR amplification products of captured templates, and longer third-generation sequencing libraries are more conducive to analyzing sequence information in pathogenic repeat regions, especially for detecting samples with very large abnormal repeat expansions.

[0011] The inventors discovered that because the lengths of randomly fragmented DNA vary, short fragments have a greater amplification advantage when amplifying long fragments using the captured template. Those skilled in the art will recognize that the amount of PCR product is related to both primer concentration and cycle number. The library construction process of the present invention does not include pre-capture PCR, which not only reduces experimental steps—eliminating the need for purification and pre-capture amplification—but also avoids amplification bias in pre-capture PCR products.

[0012] The inventors have found that by using a Y-type linker and without pre-capture PCR, the blocking sequence for blocking the linker can be omitted during hybridization without significantly affecting the capture efficiency, coverage, and coverage uniformity, thereby saving hybridization capture costs.

[0013] Therefore, in a first aspect, the present invention provides a probe set for detecting dynamically mutated STR and VNTR gene loci, targeting the following loci:

[0014] 1) STR loci (69): ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7, ATXN8OS / ATXN8, ATXN10, PPP2R2B, TBP, BEAN1 / TK2, NOP56, DAB1, ATN1, FXN, GLS, RFC1, AR, DMPK, CNBP, LRP12, GIPC1, NUTM2B-AS1 / LOC642361, PABPN1, C9orf72, NIPA1, TAF1, DMD, VWA1, SPAST, CSTB, SAMD1 2. STARD7, MARCHF6, YEATS2, TNRC6A, RAPGEF2, NOTCH2NLC, FMR1, AFF2, AFF3, ZNF713, C11ORF80, DIP2B, CBL, HTT, JPH3, PRDM12, PR NP, SOX3, ARX, ZIC2, HOXD13, EIF4A3, RUNX2, HOXA13, FOXL2, ZIC3, TBX1, XYLT1, COMP, TCF4, PHOX2B, FGF14, RILPL1, THAP11, POLG;

[0015] 2) VNTR gene loci (4): CACNA1C, PLIN4, ABCA7, and WDR7.

[0016] Furthermore, the probe set meets the following requirements:

[0017] 1) Covering 2 kb of each of the 5' and 3' flanking regions of the STR and VNTR loci, probes were designed using a 1× tiled coverage approach. For regions not suitable for probe design, such as those with poor sequence specificity, probes were not designed. When probe coverage was limited, the probe density was increased according to the following principles:

[0018] If the bases covered by the probes in the 2kb regions on both sides are between 700-1000bp, the probes are designed using a 2× tiling coverage method; if the bases covered by the probes in the 2kb regions on both sides are between 500-700bp, the probes are designed using a 3× tiling coverage method; if the bases covered by the probes in the 2kb regions on both sides are <500bp, the probes are designed using a 4× tiling coverage method.

[0019] 2) The probe length should be 80–120 bp, and the GC content should preferably be 40%–60%.

[0020] In a specific embodiment, when designing STR gene locus probes using a 4× tiling coverage method, if a probe can be designed in the target repeat region of the gene locus, it is preferred to design the probe using a 1× tiling coverage method; or, the probe can be designed by extending the 1 kb region upstream and downstream of the 5' flanking region and the 3' flanking region, that is, the probe can be designed using a 1× tiling coverage method in the 2nd to 3 kb region of the 5' flanking region and the 3' flanking region.

[0021] In a specific embodiment, for VNTR gene loci, regardless of whether the probe density is increased, if probes can be designed in the target repeat region of the gene locus, it is preferred to design probes using a 1× tiled coverage method; or extend 1kb regions upstream and downstream of its 5' flanking region and 3' flanking region to design probes, that is, design probes using a 1× tiled coverage method in the 2nd-3kb regions of its 5' flanking region and 3' flanking region.

[0022] In a specific embodiment, the probe set includes all or at least one of the following 2230 sequences:

[0023] 1) The 1× tiling probe design region includes the following gene loci:

[0024] STR gene loci: ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7, ATXN8OS / ATXN8, ATXN10, PPP2R2B, TBP, BEAN1 / TK2, NOP56, DAB1, ATN1, FXN, GLS, RFC1, AR, DMPK, CNBP, LRP12, GIPC1, PABPN1, C9orf72, NIPA1, TAF1, DMD, VWA1, CSTB, SAMD12, MARCHF6, YEATS2, TNRC6A, RAPGEF2, NOTCH2NLC, FMR1, AFF2, AFF3, ZNF713, C11ORF80, DIP2B, CBL, HTT, JPH3, PRDM12, PRNP, SOX3, ARX, ZIC2, HOXD13, EIF4A3, RUNX2, HOXA13, FOXL2, ZIC3, TBX1, XYLT1, COMP, TCF4, PHOX2B, FGF14, RILPL1, THAP11, POLG;

[0025] VNTR gene loci: CACNA1C, PLIN4, ABCA7, WDR7;

[0026] 1× tiling probe specific probe sequence (a total of 1867):

[0027] chr1:1433798-1433918;chr1:1433915-1434035;chr1:1434033-1434153;chr1:1434150-1434270;chr1:1434271-1434391;chr1:1434388-1434508;chr1:1434505-1434625;chr1:1434622-1434742;chr1:1434743-1434863;chr1:1434860-1434980;chr1:1434978-1435098;chr1:1435095-1435215;chr1:1435216-1435336;chr1:1435333-1435453;chr1:1435451-1435571;chr1:1435568-1435688;chr1:1435689-1435809;chr1:1435806-1435926;chr1:1435924-1436044;chr1:1436041-1436161;chr1:1436162-1436282;chr1:1436279-1436399;chr1:1436396-1436516;chr1:1436513-1436633;chr1:1436634-1436754;chr1:1436751-1436871;chr1:1436869-1436989;chr1:1436986-1437106;chr1:1437107-1437227;chr1:1437224-1437344;chr1:1437342-1437462;chr1:1437459-1437579;chr1:1437580-1437700;chr1:1437698-1437818;chr1:57365392-57365512;chr1:57365513-57365633;chr1:57365629-57365749;chr1:57365746-57365866;chr1:57365862-57365982;chr1:57365983-57366103;chr1:57366099-57366219;chr1:57366216-57366336;chr1:57366332-57366452;chr1:57366453-57366573;chr1:57366569-57366689;chr1:57366686-57366806;chr1:57366802-57366922;chr1:57367241-57367361;chr1:57367358-57367478;chr1:57367474-57367594;chr1:57367595-57367715;chr1:57367711-57367831;chr1:57367828-57367948;chr1:57367944-57368064;chr1:57368065-57368185;chr1:57368181-57368301;chr1:57368768-57368888;chr1:57368884-57369004;chr1:57369005-57369125;chr1:149390842-149390962;chr1:149391545-149391665;chr1:149391661-149391781;chr1:149391782-149391902;chr1:149391898-149392018;chr1:149392015-149392135;chr1:149392131-149392251;chr1:149392252-149392372;chr1:149392368-149392488;chr1:149392485-149392605;chr1:149392601-149392721;chr1:149392722-1493 92842;chr11:66742937-66743057;chr11:66743055-66743175;chr11:66743172-66743292;chr11:66743293-66743413;chr11:66743411-66743531;chr11:66743528-66743648;chr11:66743646-66743766;chr11:66743767-66743887;chr11:66744119-66744239;chr11:66744240-66744360;chr11:66744357-66744477;chr11:66744475-66744595;chr11:66744592-66744712;chr11:66744713-66744833;chr11:66744831-66744951;chr11:66744948-66745068;chr11:66745066-66745186;chr11:66745187-66745307;chr11:66745304-66745424;chr11:66745422-66745542;chr11:66745539-66745659;chr11:66745660-66745780;chr11:66746133-66746253;chr11:119204643-119204763;chr11:119204764-119204884;chr11:119204882-119205002;chr11:119205000-119205120;chr11:119205118-119205238;chr11:119205239-119205359;chr11:119205357-119205477;chr11:119205474-119205594;chr11:119205592-119205712;chr11:119205713-119205833;chr11:119205831-119205951;chr11:119205948-119206068;chr11:119206066-119206186;chr11:119206187-119206307;chr11:119206305-119206425;chr11:119206423-119206543;chr11:119206541-119206661;chr11:119206662-119206782;chr11:119206780-119206900;chr11:119206897-119207017;chr11:119207015-119207135;chr11:119207136-119207256;chr11:119207254-119207374;chr11:119207371-119207491;chr11:119207489-119207609;chr11:119207610-119207730;chr11:119207728-119207848;chr11:119207845-119207965;chr11:119207963-119208083;chr11:119208084-119208204;chr11:119208203-119208323;chr12:2253791-2253911;chr12:2253907-2254027;chr12:2254024-2254144;chr12:2254140-2254260;chr12:2254261-2254381;chr12:2254377-2254497;chr12:2254494-2254614;chr12:2254610-2254730;chr12:2254731-2254851;chr12:2254847-2254967;chr12:2254964-2255084;chr12:2255080-2255200;chr12:2255201-2255321;chr12:2255317-2255437;chr12:2255434-2255554;chr12:2255550-2255670;chr12:2255671-2255791;chr12:2256090-2256210;chr12:2256206-2256326;chr12:2256323-2256443;chr12:2256439-2256559;chr12:2256560-2256680;chr12:2256676-2256796;chr12:2256793-2256913;chr12:2256909-2257029;chr12:225 7030-2257150;chr12:2257146-2257266;chr12:2257263-2257383;chr12:2257616-2257736;chr12:2257733-2257853;chr12:2257849-2257969;chr12:2257970-2258090;chr12:2255791-2255911;chr12:2255880-2256000;chr12:2255970-2256090;chr12:6934716-6934836;chr12:6935193-6935313;chr12:6935313-6935433;chr12:6935432-6935552;chr12:6935552-6935672;chr12:6935671-6935791;chr12:6935790-6935910;chr12:6935910-6936030;chr12:6936029-6936149;chr12:6936148-6936268;chr12:6936268-6936388;chr12:6936387-6936507;chr12:6936506-6936626;chr12:6936626-6936746;chr12:6936745-6936865;chr12:6936865-6936985;chr12:6936984-6937104;chr12:6937103-6937223;chr12:6937223-6937343;chr12:6937342-6937462;chr12:6937461-6937581;chr12:6937581-6937701;chr12:6937700-6937820;chr12:6937819-6937939;chr12:6937939-6938059;chr12:6938058-6938178;chr12:6938178-6938298;chr12:6938297-6938417;chr12:6938416-6938536;chr12:6938536-6938656;chr12:6938655-6938775;chr12:50505053-50505173;chr12:50505169-50505289;chr12:50505286-50505406;chr12:50505402-50505522;chr12:50505523-50505643;chr12:50505639-50505759;chr12:50505756-50505876;chr12:50505872-50505992;chr12:50505993-50506113;chr12:50506109-50506229;chr12:50506226-50506346;chr12:50506342-50506462;chr12:50506463-50506583;chr12:50506579-50506699;chr12:50506696-50506816;chr12:50506812-50506932;chr12:50506933-50507053;chr12:111596949-111597069;chr12:111597065-111597185;chr12:111597182-111597302;chr12:111597298-111597418;chr12:111597419-111597539;chr12:111597535-111597655;chr12:111597652-111597772;chr12:111597768-111597888;chr12:111597889-111598009;chr12:111598005-111598125;chr12:111598122-111598242;chr12:111598238-111598358;chr12:111598359-111598479;chr12:111598475-111598595;chr12:111598592-111598712;chr12:111598708-111598828;chr12:111598829-111598949;chr12:111599019-111599139;chr12:111599135-111599255;chr12:111599252-111599372;chr12:111599368-111599488;chr12:111599489-111599609;chr12:111599605-111599725;chr12:111599722-11159 9842;chr12:111599838-111599958;chr12:111599959-111600079;chr12:111600075-111600195;chr12:111600192-111600312;chr12:111600308-111600428;chr12:111600429-111600549;chr12:111600545-111600665;chr12:111600662-111600782;chr13:70137383-70137503;chr13:70137501-70137621;chr13:70137620-70137740;chr13:70138214-70138334;chr13:70138453-70138573;chr13:70138572-70138692;chr13:70138690-70138810;chr13:70138811-70138931;chr13:70138929-70139049;chr13:70139048-70139168;chr13:70139166-70139286;chr13:70139287-70139407;<h2 style=";text-align:left;direction:ltr">chr13:70139523-70139643;chr13:70139641-70139761;chr13:7013976 2-70139882;chr13:70139880-70140000;chr13:70139999-70140119;chr 13:70140117-70140237;chr13:70140238-70140358;chr13:70140356-70140476;chr13:70140475-70140595;chr13:70140593-70140713;chr13: 70140714-70140834;chr13:70140832-70140952;chr13:70140951-70141071;chr13:70141069-70141189;chr13:70141190-70141310;chr13:701 41309-70141429;chr13:99983448-99983568;chr13:99983566-99983686;chr13:99983685-99983805;chr13:99983803-99983923;chr13:999839 24-99984044;chr13:99984042-99984162;chr13:99984161-99984281;chr13:99984279-99984399;chr13:99984400-99984520;chr13:99984518- 99984638;chr13:99984637-99984757;chr13:99984755-99984875;chr13:99984876-99984996;chr13:99984994-99985114;chr13:99985113-999 85233;chr13:99985231-99985351;chr13:99985352-99985472;chr13:99985470-99985590;chr13:99985588-99985708;chr13:99985706-999858 26;chr13:99985945-99986065;chr13:99986064-99986184;chr13:99986182-99986302;chr13:99986303-99986423;chr13:99986421-99986541;chr13:99986540-99986660; chr13:99986658-99986778; chr13:99986779-99986899; chr13:99986897-99987017; chr13:999870 16-99987136;chr13:99987134-99987254;chr13:99987255-99987375;chr13:99987374-99987494;chr14:23319463-23319583; chr14:23319579-23319699;chr14:23319696-23319816;chr14:23319812-23319932;chr14:23319933-23320053;chr14:23320049-23320169;chr14:23320166-23320286;chr14:23320282-23320402;chr14:23320403-23320523;chr14:23320519-23320639; chr14:23320636-23320756; chr14:23320752-23320872; chr14:23320873-23320993; chr14:23320989-23321109; chr14:23321106-23321226; chr14:23321222-23320993 21342;chr14:23321343-23321463;chr14:23321543-23321663;chr14:23321659-23321779;chr14:23321776-23321896;chr14:23321892-23322012;chr14:233220 13-23322133;chr14:23322129-23322249;chr14:23322246-23322366;chr14:23322362-23322482;chr14:23322483-23322603;chr14:23322599-23322719;chr14: 23322716-23322836;chr14:23322832-23322952;chr14:23322953-23323073;chr14:23323069-23323189;chr14:23323186-23323306;chr14:23323302-23323422;chr14:23323423-23323543;chr14:92071052-92071172;chr14:9207116 8-92071288;chr14:92071638-92071758;chr14:92071755-92071875;chr 14:92071871-92071991;chr14:92071992-92072112;chr14:92072108-9 2072228;chr14:92072225-92072345;chr14:92072341-92072461;chr14: 92072462-92072582;chr14:92072811-92072931;chr14:92072932-9207 3052;chr15:22786703-22786823;chr15:22786819-22786939;chr15:227 86936-22787056;chr15:22787052-22787172;chr15:22787173-2278729 3;chr15:22787289-22787409;chr15:22787406-22787526;chr15:227875 22-22787642;chr15:22787643-22787763;chr15:22787759-22787879;c hr15:22787876-22787996;chr15:22787992-22788112;chr15:22788113- 22788233;chr15:22788583-22788703;chr16:17468869-17468989;chr16:17468985-17469105;chr16:17469102-17469222;chr16:17469218-174 69338;chr16:17469339-17469459;chr16:17469455-17469575;chr16:17469572-17469692;chr16:17469688-17469808;chr16:17469809-174699 29;chr16:17469925-17470045;chr16:17470042-17470162;chr16:17470158-17470278;chr16:17470279-17470399;chr16:17470395-17470515;chr16:17470512-17470632;chr16:17470628-17470748;chr16:17470749-17470869;chr16:17471083-17471203;chr16:17471200-17471320;chr16:17471316-17471436;chr16:17471437-17471557;chr16:17471553-174716 73;chr16:17471670-17471790;chr16:17471786-17471906;chr16:17471907-17472027;chr16:17472023-17472143;chr16:17472140-17472260;chr16:17472256-17472376;chr16:17472847-17472967;chr16:24611438-2461 1558;chr16:24611554-24611674;chr16:24611671-24611791;chr16:24612141-2461226 1;chr16:24612257-24612377;chr16:24612378-24612498;chr16:24612727-24612847;ch r16:24612848-24612968;chr16:24613197-24613317;chr16:24613318-24613438;chr16:24613881-24614001;chr16:24614002-24614122;chr16:24614118-24614238;chr16:246 14235-24614355;chr16:24614351-24614471;chr16:24614472-24614592;chr16:24614588-24614708;chr16:24614705-24614825;chr16:24615058-24615178;chr16:24615175-2 4615295;chr16:24615291-24615411;chr16:24615412-24615532;chr16:66487396-66487516;chr16:66487514-66487634;chr16:66487633-66487753;chr16:66487751-66487871;chr16:66487872-66487992;chr16:66487990-66488110;chr16:6648810 9-66488229;chr16:66488227-66488347;chr16:66489181-66489301;chr 16:66489299-66489419;chr16:66489420-66489540;chr16:66489538-66489658;chr16:66489657-66489777;chr16:66489775-66489895;chr16: 66489896-66490016;chr16:66490014-66490134;chr16:66490490-66490610;chr16:66490609-66490729;chr16:66490727-66490847;chr16:664 90848-66490968;chr16:66490966-66491086;chr16:66491085-66491205;chr16:66491203-66491323;chr16:66491324-66491444;chr16:664914 42-66491562;chr16:66491920-66492040;chr16:66492275-66492395;chr16:66492751-66492871;chr16:66492872-66492992;chr16:66492990- 66493110;chr16:66493109-66493229;chr16:66493227-66493347;chr16:66493348-66493468;chr16:87602282-87602402;chr16:87602637-876 02757;chr16:87602758-87602878;chr16:87602876-87602996;chr16:87602995-87603115;chr16:87603113-87603233;chr16:87603234-876033 54;chr16:87603352-87603472;chr16:87603471-87603591;chr16:87603589-87603709;chr16:87603710-87603830;chr16:87603828-87603948;chr16:87603947-87604067;chr16:87604065-87604185;chr16:87604186-87604306;chr16:87604304-87604424;chr16:87604423-87604543;chr16:87604541-87604661;chr16:87604662-87604782;chr16:87604780-87604900;chr16:87604899-87605019;chr16:87605017-87605137;chr16:87605138-87605258;chr16:87605256-87605376;chr16:87605375-87605495;chr16:87605493- 87605613;chr16:87605614-87605734;chr16:87605732-87605852;chr16:87605851-87605971;chr16:87605969-87606089;chr16:87606090-87606210;chr16:87606209-87606329;chr17:80145001-80145121;chr17:80145117-80145237;chr17:80145587-80145707;chr17:80145704-80145824;chr17:80145820-80145940;chr17:80145941-80146061;chr17:80146057-80146177;chr17:80146174-80146294;chr17:80146290-80146410;chr17:80146411-80146531;chr17:80146527-80146647;chr17:80146644-80146764;chr17:80146760-80146880;chr17:80146881-80147001;chr18:55584113-55584233;chr18:55584229-55584349;chr18:55584346-55584466;chr18:55584462-55584582;chr18:55584583-55584703;chr18:55584699-55584819;chr18:55584816-55584936;chr18:55584932-55585052;chr18:55585053-55585173;chr18:55585169-55585289;chr18:55585286-55585406;chr18:55585402-55585522;chr18:55585523-55585643;chr18:55585639-55585759;chr18:55585756-55585876;chr18:55585872-55585992;chr18:55585993-55586113;chr18:55586229-55586349;chr18:55586345-55586465;chr18:55586462-55586582;chr18:55586578-55586698;chr18:55586699-55586819;chr18:55586815-55586935;chr18:55586932-55587052;chr18:55587048-55587168;chr18:55587402-55587522;chr18:55587518-55587638;chr18:55587639-55587759;chr18:55587755-55587875;chr18:55587872-55587992;chr18:55587988-55588108;chr18:55588109-55588229;chr18:57022495-57022615;chr18:57022611-57022731;chr18:57022728-57022848;chr18:57022844-57022964;chr18:57022965-57023085;chr18:57023081-57023201;chr18:57023198-57023318;chr18:57023314-57023434;chr18:57023435-57023555;chr18:57023551-57023671;chr18:57023668-57023788;chr18:57023784-57023904;chr18:57023905-57024025;chr18:57024021-57024141;chr18:57024138-57024258;chr18:57024254-57024374;<h2 style=";text-align:left;direction:ltr">chr18:57024375-57024495;chr18:57024955-57025075;chr18:57025071-57025 191;chr18:57025188-57025308;chr18:57025304-57025424;chr18:57025425-57 025545;chr18:57025541-57025661;chr18:57025658-57025778;chr18:5702577 4-57025894;chr18:57025895-57026015;chr18:57026011-57026131;chr18:5702 6128-57026248;chr18:57026244-57026364;chr18:57026365-57026485;chr18:57026481-57026601;chr 18:57026598-57026718;chr18:57026714-57026834;chr18:57026835-57026955;chr18:57024494-57024 614;chr18:57024572-57024692;chr18:57024650-57024770;chr19:1047438-1047558;chr19:1047554-1047674;chr19:1047671-1047791;chr19:1047787-1047907;chr19:1048848-1048968;chr19:1048964-104 9084;chr19:1049081-1049201;chr19:1049197-1049317;chr19:1049318-1049438;chr19:1050841-1050961;chr19:1050962-1051082;chr19:1051078-1051198;chr19:1051195-1051315;chr19:1051311-10514 31;chr19:1051432-1051552;chr19:1051548-1051668;chr19:1051665-1051785;chr19:1051781-1051901;chr19:1051902-1052022;chr19:1046437-1046557;chr19:1046543-1046663;chr19:1046650-1046770;<h2 style=";text-align:left;direction:ltr">chr19:1046756-1046876;chr19:1046877-1046997;chr19:1046983-1047103;chr19:1047090-1047210;chr19:1047196-1047316;chr19:1047317 -1047437;chr19:1052023-1052143;chr19:1052129-1052249;chr19:1052236-1052356;chr19:1052782-1052902;chr19:13206333-13206453;chr 19:13206451-13206571;chr19:13206569-13206689;chr19:13206687-13206807;chr19:13206808-13206928;chr19:13206926-13207046;chr19: 13207044-13207164;chr19:13207162-13207282;chr19:13207283-13207403;chr19:13207401-13207521;chr19:13207520-13207640;chr19:1320 7638-13207758;chr19:13207759-13207879;chr19:13207877-13207997;chr19:13207995-13208115;chr19:13208234-13208354;chr19:1320835 2-13208472;chr19:13208470-13208590;chr19:13208588-13208708;chr19:13208709-13208829;chr19:13208827-13208947;chr19:13208945-13 209065;chr19:13209063-13209183;chr19:13209184-13209304;chr19:13209302-13209422;chr19:13209420-13209540;chr19:13209538-13209 658;chr19:13209659-13209779;chr19:13209778-13209898;chr19:14494396-14494516;chr19:14494635-14494755;chr19:14494753-14494873;<h2 style=";text-align:left;direction:ltr">chr19:14494871-14494991;chr19:14494992-14495112;chr19:14495110-14495230;chr19:14495229-14495349;chr19:14495347-14495467;chr19:14495468-14495588;chr19:14495586-14495706;chr19:14495705-14495825;chr19:14495823-1449594 3;chr19:14495944-14496064;chr19:14496062-14496182;chr19:14496180-14496300;chr19:14496298-14496418;chr19:14496419-14496539;chr19:14496537-14496657;chr19:14496656-14496776;chr19:14496774-1449689 4;chr19:14496895-14497015;chr19:14497013-14497133;chr19:14497131-14497251;chr19:14497249-14497369;chr19:14497370-14497490;chr19:14497488-14497608;chr19:14497607-14497727;chr19:14497725-14497845 ;chr19:18784027-18784147;chr19:18784144-18784264;chr19:18784262-18784382;chr19:18784379-18784499;chr19:18784500-18784620;chr19:18784617-18784737;chr19:18784735-18784855;chr19:18784852-18784972 ;chr19:18784973-18785093;chr19:18785090-18785210;chr19:18785208-18785328;chr19:18785325-18785445;chr19:18785446-18785566;chr19:18785563-18785683;chr19:18785681-18785801;chr19:18785798-18785918;<h2 style=";text-align:left;direction:ltr">chr19:18785919-18786039;chr19:18786036-18786156;chr19:18786154-18786274;chr19:18786271-18786391;chr19:18786392-18786512;chr 19:18786510-18786630;chr19:18786627-18786747;chr19:18787101-18787221;chr19:18787218-18787338;chr19:18787339-18787459;chr19: 18787456-18787576;chr19:18787574-18787694;chr19:18787691-18787811;chr19:45768204-45768324;chr19:45768320-45768440;chr19:457 68437-45768557;chr19:45768553-45768673;chr19:45768674-45768794;chr19:45768790-45768910;chr19:45768907-45769027;chr19:457690 23-45769143;chr19:45769144-45769264;chr19:45769260-45769380;chr19:45769377-45769497;chr19:45769493-45769613;chr19:45769614- 45769734;chr19:45769730-45769850;chr19:45769847-45769967;chr19:45769963-45770083;chr19:45770084-45770204;chr19:45770266-457 70386;chr19:45770382-45770502;chr19:45770499-45770619;chr19:45770615-45770735;chr19:45770736-45770856;chr19:45770852-457709 72;chr19:45770969-45771089;chr19:45771085-45771205;chr19:45771206-45771326;chr19:45771322-45771442;chr19:45771439-45771559;<h2 style=";text-align:left;direction:ltr">chr19:45771555-45771675;chr19:45771676-45771796;chr19:45771792-45771912;chr19:45771909-45772029;chr19:45772025-45772145;chr19:45772146-45772266;chr2:100102799-1 00102919;chr2:100102916-100103036;chr2:100103034-100103154;chr2:100103151-100103271;chr2:100103272-10010 3392;chr2:100103390-100103510;chr2:100103507-100103627;chr2:100103625-100103745;chr2:100103746-100103866 ;chr2:100103863-100103983;chr2:100103981-100104101;chr2:100104098-100104218;chr2:100104219-100104339;chr 2:100104336-100104456;chr2:100104454-100104574;chr2:100104571-100104691;chr2:100104810-100104930;chr2:100 104927-100105047;chr2:100105045-100105165;chr2:100105166-100105286;chr2:100105283-100105403;chr2:1001054 01-100105521;chr2:100105518-100105638;chr2:100105639-100105759;chr2:100105756-100105876;chr2:100105874-1 00105994;chr2:100105991-100106111;chr2:100106112-100106232;chr2:100106230-100106350;chr2:100106347-10010 6467;chr2:100106465-100106585;chr2:100106586-100106706;chr2:100106704-100106824;chr2:176091057-176091177;chr2:176091175-176091295;chr2:176091293-176091413;chr2:176091411-176091531;chr2:176091532-176091652;chr2:176091650-176091770;chr2:176091769-176091889;chr2:176091887-176092007;chr2:176092008-176092128;chr2:176092126-176092246;chr2:176092244-176092364;chr2:176092362-176092482;chr2:176092483-176092603;chr2:176092601-176092721;chr2:176092720-176092840;chr2:176092838-176092958;chr2:176092959-176093079;chr2:176093077-176093197;chr2:176093195-176093315;chr2:176093313-176093433;chr2:176093434-176093554;chr2:176093552-176093672;chr2:176093670-176093790;chr2:176093788-176093908;chr2:176093909-176094029;chr2:176094027-176094147;chr2:176094146-176094266;chr2:176094264-176094384;chr2:176094385-176094505;chr2:176094503-176094623;chr2:176094621-176094741;chr2:176094739-176094859;chr2:176094860-176094980;chr2:176094979-176095099;chr2:190878872-190878992;chr2:190878990-190879110;chr2:190879109-190879229;chr2:190879227-190879347;chr2:190879348-190879468;chr2:190879703-190879823;chr2:190879824-190879944;<h2 style=";text-align:left;direction:ltr">chr2:190879942-190880062;chr2:190880061-190880181;chr2:190880179-190880299; chr2:190880300-190880420;chr2:190880418-190880538;chr2:19 0880537-190880657;chr2:190880655-190880775;chr2:190880776- 190880896;chr2:190880896-190881016;chr2:190881014-19088113 4;chr2:190881133-190881253;chr2:190881251-190881371;chr2:1 90881372-190881492;chr2:190881490-190881610;chr2:19088160 9-190881729;chr2:190881727-190881847;chr2:190881848-190881 968;chr2:190881966-190882086;chr2:190882085-190882205;chr2 :190882203-190882323;chr2:190882324-190882444;chr2:1908824 42-190882562;chr2:190882561-190882681;chr2:190882679-190882799;chr2:190882800-190882920;chr20:2650732-2650852;chr20:2651208-2651328;chr20:2651326-2651446;chr20:2651444-2651564;chr20:2651562-2651682;chr20:2651683-2651803;chr20:26518 01-2651921;chr20:2651920-2652040;chr20:2652038-2652158;chr20:2652159-2652279;chr20:2652277-2652397;chr20:2652395-2652515;chr20:2652513-2652633;chr20:2652634-2652754;chr20:2652752-2652872;chr20:2652871-2652991;chr20:2652989-2653109;<h2 style=";text-align:left;direction:ltr">chr20:2653110-2653230;chr20:2653228-2653348;chr20:2653346-2653466;chr20:2653464-2653584;chr20:2653585-2653705;chr20:2653940 -2654060;chr20:2654061-2654181;chr20:2654179-2654299;chr20:2654297-2654417;chr20:2654415-2654535;chr20:2654536-2654656;chr20 :2654655-2654775;chr20:4697398-4697518;chr20:4697516-4697636;chr20:4697635-4697755;chr20:4697753-4697873;chr20:4697874-4697 994;chr20:4697992-4698112;chr20:4698111-4698231;chr20:4698229-4698349;chr20:4698350-4698470;chr20:4698587-4698707;chr20:4698 705-4698825;chr20:4698826-4698946;chr20:4698944-4699064;chr20 :4699063-4699183;chr20:4699181-4699301;chr20:4699302-4699422;c hr20:4699422-4699542;chr20:4699540-4699660;chr20:4699659-4699779;chr20:4699777-4699897;chr20:4699898-4700018;chr20:4700016-4 700136;chr20:4700135-4700255;chr20:4700253-4700373;chr20:4700374-4700494;chr20:4700492-4700612;chr20:4700611-4700731;chr20: 4700729-4700849;chr20:4700850-4700970;chr20:4700968-4701088;chr20:4701087-4701207;chr20:4701205-4701325;chr20:4701326-470144 6;<h2 style=";text-align:left;direction:ltr">chr21:43774444-43774564;chr21:43774560-43774680;chr21:4377467 7-43774797;chr21:43774793-43774913;chr21:43775263-43775383;chr 21:43775384-43775504;chr21:43775500-43775620;chr21:43775617-4 3775737;chr21:43775733-43775853;chr21:43775854-43775974;chr21: 43775970-43776090;chr21:43776087-43776207;chr21:43776203-43776323;chr21:43776324-43776444;chr22:19764762-19764882;chr22:197 64880-19765000;chr22:19764999-19765119;chr22:19765117-19765237;chr22:19765238-19765358;chr22:19765356-19765476;chr22:197654 75-19765595;chr22:19765593-19765713;chr22:19765714-19765834;chr22:19765832-19765952;chr22:19765950-19766070;chr22:19766068- 19766188;chr22:19766189-19766309;chr22:19766307-19766427;chr22:19766426-19766546;chr22:19766544-19766664;chr22:19766665-197 66785;chr22:19766783-19766903;chr22:19766902-19767022;chr22:19767020-19767140;chr22:19767141-19767261;chr22:19767259-197673 79;chr22:19767378-19767498;chr22:19767496-19767616;chr22:19767617-19767737;chr22:19767735-19767855;chr22:19767853-19767973;<h2 style=";text-align:left;direction:ltr">chr22:19767971-19768091;chr22:19768092-19768212;chr22:19768210-19768330;chr22:19768329-19768449;chr22:19768447-19768567;chr22:19768568-19768688;chr22:19768687-19768807;chr22:45793354-45793474;chr22:45793470-45793590;chr22:45793587-45 793707;chr22:45793703-45793823;chr22:45793824-45793944;chr22:45794173-45794293;chr22:45794410-45794530;chr22:45794527-45794647;chr22:45794643-45794763;chr22:45794764-45794884;chr22:45794880-45795000;chr22:45794997-45795117;chr22:4579 5113-45795233;chr22:45795234-45795354;chr22:45795330-45795450;chr3:63910684-63910804;chr3:63910802-63910922;chr3:63910919-63911039;chr3:63911037-63911157;chr3:63911158-63911278;chr3:63911276-63911396;chr3:63911393-63911513;chr3:63911 511-63911631;chr3:63911632-63911752;chr3:63911750-63911870 ;chr3:63911868-63911988;chr3:63911986-63912106;chr3:639121 07-63912227;chr3:63912225-63912345;chr3:63912342-63912462; chr3:63912460-63912580;chr3:63912581-63912701;chr3:63912699 -63912819;chr3:63912816-63912936;chr3:63912934-63913054;chr3:63913055-63913175;chr3:63913173-63913293;chr3:63913290-63913410;chr3:63913408-63913528;chr3:63913529-63913649;chr3:63913647-63913767;chr3:63913764-63913884;chr3:63913882-63914002;chr3:63914003-63914123;chr3:63914121-63914241;chr3:63914238-63914358;chr3:63914356-63914476;chr3:63914477-63914597;chr3:63914596-63914716;chr3:129170576-129170696;chr3:129170692-129170812;chr3:129170809-129170929;chr3:129170925-129171045;chr3:129171046-129171166;chr3:129171162-129171282;chr3:129171279-129171399;chr3:129171395-129171515;chr3:129171516-129171636;chr3:129171632-129171752;chr3:129171749-129171869;chr3:129172219-129172339;chr3:129172775-129172895;chr3:129172892-129173012;chr3:129173008-129173128;chr3:129173245-129173365;chr3:129173362-129173482;chr3:129173478-129173598;chr3:129173599-129173719;chr3:129173715-129173835;chr3:129173832-129173952;chr3:129173948-129174068;chr3:129174069-129174189;chr3:129174185-129174305;chr3:138944021-138944141;chr3:138944139-138944259;chr3:138944257-138944377;chr3:138944375-138944495;chr3:138944496-138944616;chr3:138944614-138944734;chr3:138944733-138944853;chr3:138944851-138944971;chr3:138944972-138945092;chr3:138945090-138945210;chr3:138945208-138945328;chr3:138945326-138945446;chr3:138945447-138945567;chr3:138945565-138945685;chr3:138945683-138945803;chr3:138945801-138945921;chr3:138945922-138946042;chr3:138946040-138946160;chr3:138946158-138946278;chr3:138946276-138946396;chr3:138946397-138946517;chr3:138946515-138946635;chr3:138946634-138946754;chr3:138946752-138946872;chr3:138946873-138946993;chr3:138946991-138947111;chr3:138947109-138947229;chr3:138947227-138947347;chr3:138947348-138947468;chr3:138947466-138947586;chr3:138947584-138947704;chr3:138947702-138947822;chr3:138947823-138947943;chr3:138947942-138948062;chr3:183710176-183710296;chr3:183710292-183710412;chr3:183710409-183710529;chr3: 183710525-183710645;chr3:183710646-183710766;chr3:183710762-183710882;chr3:183710879-183710999;chr3:183710995-183711115;chr3:183711116-183711236;chr3:183711465-183711585;chr3:183711586-183711706;chr3:183711702-183711822;chr4:3073109-3073229;chr4:3073225-3073345;chr4:3073346-3073466;chr4:3073462-3073582;chr4:3073579-3073699;chr4:3073695-3073815;chr4:3073816-3073936;chr4:3073932-3074052;chr4:3074049-3074169;chr4:3074165-3074285;chr4:3074286-3074406;chr4:3074402-3074522;chr4:3074519-3074639;chr4:3074635-3074755;chr4:3074756-3074876;chr4:3074940-3075060;chr4:3075056-3075176;chr4:3075173-3075293;chr4:3075289-3075409;chr4:3075410-3075530;chr4:3075526-3075646;chr4:3075643-3075763;chr4:3075759-3075879;chr4:3075880-3076000;chr4:3075996-3076116;chr4:3076113-3076233;chr4:3076229-3076349;chr4:3076350-3076470;chr4:3076466-3076586;chr4:3076583-3076703;chr4:3076699-3076819;chr4:39346543-39346663;chr4:39346663-39346783;chr4:39346782-39346902;chr4:39346901-39347021;chr4:39347021-39347141;chr4:39347140-39347260;chr4:39347260-39347380;chr4:39347379-39347499;chr4:39347618-39347738;chr4:39347737-39347857;chr4:39347856-39347976;chr4:39347976-39348096;chr4:39348453-39348573;chr4:39348573-39348693;chr4:39348692-39348812;chr4:39348811-39348931;chr4:39348931-39349051;chr4:39349050-39349170;chr4:39349289-39349409;chr4:39349408-39349528;chr4:39349527-39349647;chr4:39349647-39349767;chr4:39350124-39350244;chr4:39350244-39350364;chr4:39350363-39350483;chr4:41744093-41744213;chr4:41744212-41744332;chr4:41744330-41744450;chr4:41744451-41744571;chr4:41744569-41744689;chr4:41744688-41744808;chr4:41744806-41744926;chr4:41744927-41745047;chr4:41745045-41745165;chr4:41745164-41745284;chr4:41745403-41745523;chr4:41745521-41745641;chr4:41745640-41745760;chr4:41745758-41745878;chr4:41745879-41745999;chr4:41745997-41746117;chr4:41746116-41746236;chr4:41746234-41746354;chr4:41746355-41746475;chr4:41746473-41746593;chr4:41746592-4 1746712;chr4:41746710-41746830;chr4:41746831-41746951;chr4:41746949-41747069;chr4:41747068-41747188;chr4:41747186-41747306;chr4:41747307-41747427;chr4:41747425-41747545;chr4:41747544-41747664;chr4:41747662-41747782;chr4:41747783-41747903;chr4:41747902-41748022;chr4:159340526-159340646;chr4:159340875-159340995;chr4:159340996-159341116;chr4:159341112-159341232;chr4:159341229-159341349;chr4:159341345-159341465;chr4:159341466-159341586;chr4:159341582-159341702;chr4:159341699-159341819;chr4:159341815-159341935;chr4:159341936-159342056;chr4:159342052-159342172;chr4:159342169-159342289;chr4:159342285-159342405;chr4:159342406-159342526;chr4:159342618-159342738;chr4:159342734-159342854;chr4:159342851-159342971;chr4:159342967-159343087;chr4:159343088-159343208;chr4:159343204-159343324;chr4:159343321-159343441;chr4:159343437-159343557;chr4:159343558-159343678;chr4:159343674-159343794;chr4:159343791-159343911;chr4:159343907-159344027;chr4:159344028-159344148;chr4:159344144-159344264;chr4:159344261-159344381;chr4:159344377-159344497;chr4:159344498-159344618;chr5:10354338-10354458;chr5:10354454-10354574;chr5:10354571-10354691;chr5:10354687-10354807;chr5:10354808-10354928;<h2 style=";text-align:left;direction:ltr">chr5:10354924-10355044;chr5:10355041-10355161;chr5:10355157-10355277;chr5:10355278-10355398;chr5:10355394-10355514;chr5:10355511-10355631;chr5:10355627-10355747;chr5:10355748-10355868; chr5:10355864-10355984;chr5:10355981-10356101;chr5:10356097-10356217;chr5:10356218-10356338;chr5:10356760-10356880;chr5:10356881-10357001;chr5:10356997-10357117;chr5:10357114-10357234; chr5:10357230-10357350;chr5:10357351-10357471;chr5:10357467-10357587;chr5:10357584-10357704;chr5:10357700-10357820;chr5:10357821-10357941;chr5:10358054-10358174;chr5:10358170-10358290; chr5:10358291-10358411;chr5:146876727-146876847;chr5:146876845-146876965;chr5:146876962-146877082;chr5:146877080-146877200;chr5:146877201-146877321;chr5:146877319-146877439;chr5:1468774 36-146877556;chr5:146877554-146877674;chr5:146877675-146877795;chr5:146877793-146877913;chr5:146877911-146878031;chr5:146878029-146 878149;chr5:146878150-146878270;chr5:146878268-146878388;chr5:146878385-146878505;chr5:146878503-146878623;chr5:146878624-146878744;chr5:146878742-146878862;chr5:146878859-146878979;chr5:1468789 77-146879097;chr5:146879098-146879218;chr5:146879216-146879336 ;chr5:146879333-146879453;chr5:146879451-146879571;chr5:146879 572-146879692;chr5:146879690-146879810;chr5:146879807-146879927 ;chr5:146879925-146880045;chr5:146880046-146880166;chr5:146880281-146880401;chr5:146880399-146880519;chr5:146880520-146880640;chr5:146880639-146880759;chr6:16325633-16325753;chr6:16325982-16326102;chr6:16326103-16326223;chr6:16326219-16326339;chr6:16 326336-16326456;chr6:16326452-16326572;chr6:16326573-16326693; chr6:16326689-16326809;chr6:16326806-16326926;chr6:16326922-16 327042;chr6:16327043-16327163;chr6:16327159-16327279;chr6:1632 7276-16327396;chr6:16327392-16327512;chr6:16327513-16327633;chr 6:16327724-16327844;chr6:16327840-16327960;chr6:16327957-16328077;chr6:16328073-16328193;chr6:16328194-16328314;chr6:16328310-16328430;chr6:16328427-16328547;chr6:16328543-16328663;chr6:16329013-16329133;chr6:16329134-16329254;chr6:16329250-16329370;chr6:16329367-16329487;chr6:16329483-16329603;chr6:16329604-16329724;chr6:45420678-45420798;chr6:45420794-45420914;chr6:45420911-45421031;chr6:45421027-45421147;ch r6:45421148-45421268;chr6:45421264-45421384;chr6:45421381-45421501;chr6:45421497- 45421617;chr6:45421618-45421738;chr6:45421734-45421854;chr6:45421851-45421971;chr6 :45421967-45422087;chr6:45422088-45422208;chr6:45422204-45422324;chr6:45422321-45422441;chr6:45422437-45422557;chr6:45422558-45422678;chr6:45422801-45422921;chr6:4 5422917-45423037;chr6:45423034-45423154;chr6:45423150-45423270;chr6:45423271-45423391;chr6:45423387-45423507;chr6:45423504-45423624;chr6:45423620-45423740;chr6:454 23741-45423861;chr6:45423857-45423977;chr6:45423974-45424094;chr6:45424090-45424210;chr6:45424211-45424331;chr6:45424327-45424447;chr6:4542444-45424564;chr6:4542 4560-45424680;chr6:45424681-45424801;chr6:170562017-170562137;chr6:170562133-170562253;chr6:170562250-170562370;chr6:170562366-170562486;chr6:170562487-170562607;<h2 style=";text-align:left;direction:ltr">chr6:170562603-170562723;chr6:170562720-170562840;chr6:170562 836-170562956;chr6:170562957-170563077;chr6:170563073-17056319 3;chr6:170563190-170563310;chr6:170563306-170563426;chr6:17056 3427-170563547;chr6:170563543-170563663;chr6:170563660-1705637 80;chr6:170563776-170563896;chr6:170563897-170564017;chr7:2719 7678-27197798;chr7:27197794-27197914;chr7:27197911-27198031;ch r7:27198027-27198147;chr7:27198148-27198268;chr7:27198264-27198384;chr7:27198381-27198501;chr7:27198497-27198617;chr7:271986 18-27198738;chr7:27198734-27198854;chr7:27198851-27198971;chr7:27198967-27199087;chr7:27199088-27199208;chr7:27199204-27199 324;chr7:27199321-27199441;chr7:27199437-27199557;chr7:27199558-27199678;chr7:27199967-27200087;chr7:27200083-27200203;chr7: 27200200-27200320;chr7:27200316-27200436;chr7:27200437-27200557;chr7:27200553-27200673;chr7:27200670-27200790;chr7:27200786- 27200906;chr7:27200907-27201027;chr7:27201023-27201143;chr7:27201140-27201260;chr7:27201256-27201376;chr7:27201377-27201497;chr7:27201493-27201613;chr7:27201610-27201730;chr7:27201726-27201846;chr7:27201847-27201967;chr8:104586964-104587084;chr8:104587082-104587202;chr8:104587200-104587320;chr8:104587318-104587438;chr8:104587439-104587559;chr8:104587557-104587677;chr8:104587674-104587794;chr8:104587792-104587912;chr8:104587913-104588033;chr8:104588031-104588151;chr8:104588149-104588269;chr8:104588267-104588387;chr8:104588388-104588508;chr8:104588506-104588626;chr8:104588623-104588743;chr8:104588741-104588861;chr8:104589098-104589218;chr8:104589216-104589336;chr8:104589337-104589457;chr8:104589455-104589575;chr8:104 589572-104589692;chr8:104589690-104589810;chr8:104589811-104589931;chr8:104589929-104590049;chr8:104590047-104590167;chr8:104590165-104590285;chr8:104590286-104590406;chr8:104590760-104590880;chr8:104590879-104590999;chr8:118364812-118364932;chr8:118364928-118365048;chr8:118365045-118365165;chr8:118365161-118365281;chr8:118365398-118365518;chr8:118365515-118365635;chr8:118365752-118365872;<h2 style=";text-align:left;direction:ltr">chr8:118365868-118365988;chr8:118365985-118366105;chr8:118366 101-118366221;chr8:118366222-118366342;chr8:118366338-11836645 8;chr8:118366918-118367038;chr8:118367034-118367154;chr8:1183 67151-118367271;chr8:118367267-118367387;chr8:118367388-118367 508;chr8:118367504-118367624;chr8:118367621-118367741;chr8:11 8367737-118367857;chr8:118367858-118367978;chr8:118367974-1183 68094;chr8:118368091-118368211;chr8:118368207-118368327;chr8:1 18368328-118368448;chr8:118368444-118368564;chr8:118368561-118 368681;chr8:118368677-118368797;chr8:118368798-118368918;chr9:27571717-27571837;chr9:27571833-27571953;chr9:27571954-275720 74;chr9:27572070-27572190;chr9:27572187-27572307;chr9:27572303-27572423;chr9:27572424-27572544;chr9:27572540-27572660;chr9:2 7572657-27572777;chr9:27572773-27572893;chr9:27572894-27573014;chr9:27573010-27573130;chr9:27573127-27573247;chr9:27573243- 27573363;chr9:27573364-27573484;chr9:27573546-27573666;chr9:27573662-27573782;chr9:27573779-27573899;chr9:27573895-27574015;<h2 style=";text-align:left;direction:ltr">chr9:27574016-27574136;chr9:27574132-27574252;chr9:27574249-27574369;chr9:27574365-27574485;chr9:27574486-27574606;chr9:27574602-27574 722;chr9:27574719-27574839;chr9:27574835-27574955;chr9:27574956-27575076;chr9:27575072-27575192;chr9:27575189-27575309;chr9:27575426-27 575546;chr9:69035275-69035395;chr9:69035391-69035511;chr9:69035508-69035628;chr9:69035624-69035744;chr9:69035745-69035865;chr9:6903586 1-69035981;chr9:69035978-69036098;chr9:69036094-69036214;chr9:69036215-69036335;chr9:69036448-69036568;chr9:69036564-69036684;chr9:6903 6685-69036805;chr9:69036801-69036921;chr9:69036918-69037038;chr9:69037034-69037154;chr9:130681641-130681761;chr9:130681757-130681877;chr9:130681874-130681994;chr9:130681990-130682110;chr9:130682111-130682231;chr9:130682227-130682 347;chr9:130682344-130682464;chr9:130682460-130682580;chr9:130682581-130682701;chr9:130682697-130682817;chr9:130682814-130682934;chr9:130682930-130683050;chr9:130683051-130683171;chr9:130683167-130683287;chr9:130683284-130683404;chr9:130683400-130683520;chr9:130683521-130683641;chrX:25011529-25011649;chrX:25011645-25011765;chrX:25011762-25011882;chrX:25011878-25011998;chrX:25011999-25012119;chrX:25012115-25012235;chrX:25012232-25012352;chrX:25012348-25012468;chrX:25012469-25012589;chrX:25012585-25012705;chrX:25012702-25012822;chrX:25012818-25012938;chrX:25012939-25013059;chrX:25013055-25013175;chrX:25013172-25013292;chrX:25013288-25013408;chrX:25013409-25013529;chrX:25013697-25013817;chrX:25013813-25013933;chrX:25014283-25014403;chrX:25014400-25014520;chrX:25014516-25014636;chrX:25014637-25014757;chrX:25014753-25014873;chrX:25014870-25014990;chrX:25014986-25015106;chrX:25015107-25015227;chrX:25015223-25015343;chrX:25015340-25015460;chrX:25015456-25015576;chrX:25015577-25015697;chrX:31282557-31282677;chrX:31282675-31282795;chrX:31282794-31282914;chrX:31282912-31283032;chrX:31283033-31283153;chrX:31283152-31283272;chrX:31283272-31283392;chrX:31283390-31283510;chrX:31283509-31283629;chrX:31283627-31283747;chrX:31283748-31283868;chrX:31283866-31283986;chrX:31284343-31284463;chrX:31284462-31284582;chrX:31284700-31284820;chrX:31284819-31284939;chrX:31284937-31285057;chrX:31285058-31285178;chrX:31285176-31285296;chrX:31285295-31285415;chrX:31285413-31285533;chrX:31285534-31285654;chrX:31285653-31285773;chrX:31285772-31285892;chrX:31285892-31286012;chrX:31286010-31286130;chrX:31286129-31286249;chrX:31286247-31286367;chrX:31286368-31286488;c hrX:31286487-31286607;chrX:67543316-67543436;chrX:67543432-67543552;chrX:67543549-67543669;chrX:67543665-67543785;chrX:67543786-67543906;chrX:67543902-67544022;chrX:67544019-67544139;chrX:67544135-67544255;chrX:67544256-67544376;chrX:67544372-67544492;chrX:67544489-67544609;chrX:67544605-67544725;chrX:67544726-67544846;chrX:67544842-67544962;chrX:67544959-67545079;chrX:67545075-67545195;chrX:67545196-67545316;chrX:67545419-67545539;chrX:67545535-67545655;chrX:67545652-67545772;chrX:67545768-67545888;chrX:67545889-67546009;chrX:67546005-67546125;chrX:67546122-67546242;chrX:67546238-67546358;chrX:67546359-67546479;chrX:67546475-67546595;chrX:67546592-67546712;chrX:67546708-67546828;chrX:67546829-67546949;chrX:67546945-67547065;chrX:67547062-67547182;chrX:67547178-67547298;chrX:67547299-67547419;chrX:71449479-71449599;chrX:71449595-71449715;chrX:71450535-71450655;chrX:71450652-71450772;chrX:71450768-71450888;chrX:71450889-71451009;chrX:71451005-71451125;chrX:71451122-71451242;chrX:71451238-71451358;chrX:71451359-71451479;chrX:71453833-71453953;chrX:71453949-71454069;chrX:71454070-71454190;chrX:71454186-71454306;chrX:71454419-71454539;chrX:71454540-71454660;chrX:71454656-71454776;chrX:71454773-71454893;chrX:71454889-71455009;chrX:71455010-71455130;chrX:137564825-137564945;chrX:137564943-137565063;chrX:137565060-137565180;chrX:137565178-137565298;chrX:137565299-137565419;chrX:137565416-137565536;chrX:137565534-137565654;chrX:137565651-137565771;chrX:137565772-137565892;chrX:137565890-137566010;chrX:137566007-137566127;chrX:137566125-137566245;chrX:137566246-137566366;chrX:137566363-137566483;chrX:137566481-137566601;chrX:137566598-137566718;chrX:137566719-137566839;chrX:137566837-137566957;chrX:137566954-137567074;chrX:137567072-137567192;chrX:137567193-137567313;chrX:137567310-137567430;chrX:137567428-137567548;chrX:137567545-137567665;chrX:137567666- 137567786;chrX:137567784-137567904;chrX:137567901-137568021;chrX:137568019-137568139;chrX:137568140-137568260;chrX:137568258-137568378;chrX:137568375-137568495;chrX:137568493-137568613;chrX:137568614-137568734;chrX:137568732-137568852;chrX:140502316-140502436;chrX:140502432-140502552;chrX:140502549-140502669;chrX:140502665-140502785;chrX:140502786-140502906;chrX:140502902-140503022;chrX:140503019-140503139;chrX:140503135-140503255;chrX:140503256-140503376;chrX:140503372-140503492;chrX:140503489-140503609;chrX:140503605-140503725;chrX:140503726-140503846;chrX:140503842-140503962;chrX:140503959-140504079;chrX:140504075-140504195;chrX:140504196-140504316;chrX:140504381-140504501;chrX:140504497-140504617;chrX:140504614-140504734;chrX:140504730-140504850;chrX:140504851-140504971;chrX:140504967-140505087;chrX:140505084-140505204;chrX:140505200-140505320;chrX:140505321-140505441;chrX:140505437-140505557;chrX:140505554-140505674;chrX:140505670-140505790;chrX:140505791-140505911;chrX:140505907-140506027;chrX:140506024-140506144;chrX:140506140-140506260;chrX:140506261-140506381;chrX:147909978-147910098;chrX:147910094-147910214;chrX:147910211-147910331;chrX:147910327-147910447;chrX:147910448-147910568;chrX:147910564-147910684;chrX:147910681-147910801;chrX:147910797-147910917;chrX:147910918-147911038;chrX:147911034-147911154;chrX:147911151-147911271;chrX:147911267-147911387;chrX:147911388-147911508;chrX:147911504-147911624;chrX:147911621-147911741;chrX:147911737-147911857;chrX:147911858-147911978;chrX:147912111-147912231;chrX:147912227-147912347;chrX:147912344-147912464;chrX:147912460-147912580;chrX:147912581-147912701;chrX:147912697-147912817;chrX:147912814-147912934;chrX:147912930-147913050;chrX:147913051-147913171;chrX:147913167-147913287;chrX:147913284-147913404;chrX:147913400-147913520;chrX:147913521-147913641;chrX:147913637-147913757;chrX:147913754-14 7913874;chrX:147913870-147913990;chrX:147913991-147914111;chrX:148498601-148498721;chrX:148498717-148498837;chrX:148498834-148498954;chrX:148498950-148499070;chrX:148499071-148499191;chrX:148499187-148499307;chrX:148499304-148499424;chrX:148499420-148499540;chrX:148499541-148499661;chrX:148499657-148499777;chrX:148499774-148499894;chrX:148499890-148500010;chrX:148500011-148500131;chrX:148500127-148500247;chrX:148500244-148500364;chrX:148500360-148500480;chrX:148500481-148500601;chrX:148500743-148500863;chrX:148500859-148500979;chrX:148500976-148501096;chrX:148501092-148501212;chrX:148501213-148501333;chrX:148501329-148501449;chrX:148501446-148501566;chrX:148501562-148501682;chrX:148501683-148501803;chrX:148501799-148501919;chrX:148501916-148502036;chrX:148502032-148502152;chrX:148502153-148502273;chrX:148502269-148502389;chrX:148502386-148502506;chrX:148502502-148502622;chrX:148502623-148502743;chr13:102159577-102159697;chr13:102159695-102159815;chr13:102159812-102159932;chr13:102159930-102160050;chr13:102160051-102160171;chr13:102160169-102160289;chr13:102160286-102160406;chr13:102160404-102160524;chr13:102160525-102160645;chr13:102160643-102160763;chr13:102160760-102160880;chr13:102160878-102160998;chr13:102160999-102161119;chr13:102161117-102161237;chr13:102161234-102161354;chr13:102161352-102161472;chr13:102161473-102161593;chr13:102161708-102161828;chr13:102161826-102161946;chr13:102161947-102162067;chr13:102162065-102162185;chr13:102162182-102162302;chr13:102162300-102162420;chr13:102162421-102162541;chr13:102162539-102162659;chr13:102162656-102162776;chr13:102162774-102162894;chr13:102162895-102163015;chr13:102163013-102163133;chr13:102163130-102163250;chr13:102163248-102163368;chr13:102163369-102163489;chr13:102163488-102163608;chr13:102163606-102163726;chr15:89331580-89331700;chr15:89331698-89331818;chr15:89331817-89331937;chr15:89331935-89332055;chr1 5:89332056-89332176;chr15:89332175-89332295;chr15:89332295-89332415;chr15:89332413-89332533;chr15:89332532-89332652;chr15:89332650-89332770;chr15:89332771-89332891;chr15:89332889-89333009;chr15:89333008-89333128;chr 15:89333126-89333246;chr15:89333247-89333367;chr15:89333366-89333486;chr15:89333485-89333605;chr15:89333605-89333725;chr15:89333723-89333843;chr15:89333842-89333962;chr15:89333960-89334080;chr15:89334081-89334201;ch r15:89334199-89334319;chr15:89334318-89334438;chr15:89334436-89334556;chr15:89334557-89334677;chr15:89334676-89334796;chr15:89334795-89334915;chr15:89334915-89335035;chr15:89335033-89335153;chr15:89335152-89335272;c hr15:89335270-89335390;chr15:89335391-89335511;chr15:89335510-89335630;chr16:67841223-67841343;chr16:678413 43-67841463;chr16:67841463-67841583;chr16:67841584-67841704;chr16:67841704-67841824;chr16:67841824-67841944;chr16:67841944-67842064;chr16:67842065-67842185;chr16:67842185-67842305;chr16:67842305-67842425;chr16:67842426-67842546;chr16:67842546-67842666;chr16:67842666-67842786;chr16:67842786-67842906;chr16:67842907-67843027;chr16:67843027-67843147;chr16:67843147-67843267;chr16:67843267-67843387;chr16:67843388-67843508;chr16:67843508-67843628;chr16:67843628-67843748;chr16:67843749-67843869;chr16:67843869-67843989;chr16:67843989-67844109;chr16:67844109-67844229;chr16:67844230-67844350;chr16:67844350-67844470;chr16:67844470-67844590;chr16:67844590-67844710;chr16:67844711-67844831;chr12:123531838-123531958;chr12:123531956-123532076;chr12:123532074-123532194;chr12:123532195-123532315;chr12:123532313-123532433;chr12:123532430-123532550;chr12:123532548-123532668;chr12:123532669-123532789;chr12:123532787-123532907;chr12:123532905-123533025;chr12:123533023-123533143;chr12:123533144-123533264;chr12:123533262-123533382;chr12:123533379-123533499;chr12:123533497-123533617;chr12:123533618-123533738;<h2 style=";text-align:left;direction:ltr">chr12:123533736-123533856;chr12:123533854-123533974;chr12:12 3533972-123534092;chr12:123534093-123534213;chr12:12353421 1-123534331;chr12:123534328-123534448;chr12:123534446-1235 34566;chr12:123534567-123534687;chr12:123534685-123534805; chr19:4510446-4510566;chr19:4510561-4510681;chr19:4510675- 4510795;chr19:4510790-4510910;chr19:4510911-4511031;chr19:4511026-4511146;chr19:4511140-4511260;chr19:4511255-4511375;chr19:4511376-4511496;chr19:4511491-4511611;chr19:4511605-4511725;chr19:4511720-4511840;chr19:4511841-4511961;chr1 9:4511956-4512076;chr19:4512070-4512190;chr19:4512185-4512305;chr19:4512306-4512426;chr19:4512420-4512540;chr19:4512535-4512655;chr19:4512649-4512769;chr19:4512770-4512890;chr19:4512885-4513005;chr19:4512999-4513119;chr19:4513114-45 13234;chr19:4513235-4513355;chr19:4513352-4513472;chr19:4513468-4513588;chr19:4513584-4513704;chr7:55884919-55885039 ;chr7:55885040-55885160;chr7:55889640-55889760;chr7:55889746-55889866;chr7:55889853-55889973;chr7:55889959-55890079;chr7:55890520-55890640;

[0028] 2) The 2× tiling probe design region includes the following gene loci:

[0029] STR gene loci: TBP, FXN, NUTM2B-AS1 / LOC642361, NIPA1, SPAST, CSTB, YEATS2, NOTCH2NLC, ZNF713, PRDM12, EIF4A3;

[0030] The specific sequences of the 2× tiling probes are (a total of 217):

[0031] chr1:149388722-149388842;chr1:149388782-149388902;chr1:149388842-149388962;chr1:149388902-149389022;chr1:149388962-149389082;chr1:149389022-149389142;chr1:149389082-149389202;chr1:149389142-149389262;chr1:149389202-149389322;chr1:149389262-149389382;chr1:149389322-149389442;chr1:149389382-149389502;chr1:149389442-149389562;chr1:149389502-149389622;chr1:149389562-149389682;chr1:149389622-149389742;chr1:149389682-149389802;chr1:149389742-149389862;chr1:149389802-149389922;chr1:149389862-149389982;chr1:149390762-149390882;chr10:79824234-79824354;chr10:79824294-79824414;chr10:79824354-7 9824474;chr10:79824414-79824534;chr10:79824474-79824594;chr10:79824534-79824654;chr10:79824594-79824714;chr10:79824654-79824774;chr10:79824714-79824834;chr10:79824774-79824894;chr10:79825014-79825134;chr10:79825074-79825194;chr10:79825134-79825254;chr10:79825194-79825314;chr10:79825254-79825374;chr10:79825314-79825434;chr10:79825374-79825494;chr10:79825434-79825554;chr10:79825494-79825614;chr10:79827344-79827464;chr10:79827404-79827524;chr10:79827464-79827584;chr10:79827524-79827644;chr10:79827584-79827704;chr10:79827644-79827764;chr10:79827704-79827824;chr10:79828064-79828184;chr10:79828124-79828244;chr10:79828184-79828304;chr10:79828244-79828364;chr10:79828304-79828424;chr10:79828364-79828484;chr15:22784771-22784891;chr15:22785491-22785611;chr15:22785551-22785671;chr15:22785611-22785731;chr15:22786091-22786211;chr15:22786151-22786271;chr15:22786211-22786331;chr15:22786271-22786391;chr15:22786331-22786451;chr15:22786391-22786511;chr15:22786451-22786571;chr15:22786511-22786631;chr15:22786571-22786691;chr15:22786631-22786751;chr17:80147039-80147159;chr17:80147099-80147219;chr17:80147159-80147279;chr17:80147219-80147339;chr17:80147279-80147399;chr17:80147339-80147459;chr17:80147399-80147519;chr17:80147459-80147579;chr17:80147519-80147639;chr17:80147579-80147699;chr17:80147639-80147759;chr17:80147699-80147819;chr17:80147999-80148119;chr17:80148059-80148179;chr17:80148119-80148239;chr17:80148359-80148479;chr17:80148419-80148539;chr2:32071717-32071837;chr2:32071777-32071897;chr2:32071837-32071957;chr2:32072317-32072437;chr2:32072377-32072497;chr2:32072557-32072677;chr2:32072617-32072737;chr2:32072677-32072797;chr2:32072737-32072857;chr2:32072797-32072917;chr2:32072857-32072977;chr2:32072917-32073037;chr2:32072977-32073097;chr2:32073037-32073157;chr2:32073097-32073217;chr2:32073277-32073397;chr2:32073337-32073457;chr2:32073397-32073517;chr2:32073577-32073697;chr2:32073637-32073757;chr2:320 73697-32073817;chr2:32073757-32073877;chr2:32073817-32073937;chr2:32073877-32073997;chr2:32073937-32074057;chr2:32073997-32074117;chr2:32074057-32074177;chr2:32074117-32074237;chr2:32074177-32074297;chr2:32074237-32074357;chr2:32074297-32074417;chr2:32074357-32074477;chr2:32074417-32074537;chr2:32074777-32074897;chr2:32074837-32074957;chr2:32074897-32075017;chr2:32074957-32075077;chr2:32075017-32075137;chr21:43776399-43776519;chr21:43776459-43776579;<h2 style=";text-align:left;direction:ltr">chr21:43776519-43776639;chr21:43776579-43776699;chr21:4377663 9-43776759;chr21:43776699-43776819;chr21:43776759-43776879;chr 21:43776819-43776939;chr21:43776879-43776999;chr21:43777959-4 3778079;chr21:43778019-43778139;chr21:43778079-43778199;chr21: 43778139-43778259;chr21:43778199-43778319;chr21:43778259-4377 8379;chr21:43778319-43778439;chr21:43778379-43778499;chr21:437 78439-43778559;chr3:183712446-183712566;chr3:183712506-183712 626;chr3:183712566-183712686;chr3:183712626-183712746;chr3:183 712686-183712806;chr3:183713046-183713166;chr3:183713106-1837 13226;chr3:183713166-183713286;chr3:183713526-183713646;chr3:1 83713586-183713706;chr3:183713646-183713766;chr3:183713706-183713826;chr6:170559826-170559946;chr6:170559886-170560006;chr6 :170559946-170560066;chr6:170560006-170560126;chr6:170560966- 170561086;chr6:170561266-170561386;chr6:170561326-170561446;ch r6:170561386-170561506;chr6:170561446-170561566;chr6:170561506-170561626;chr6:170561566-170561686;chr6:170561626-170561746;<h2 style=";text-align:left;direction:ltr">chr6:170561686-170561806;chr6:170561746-170561866;chr6:170561806-170561926;chr7:55885581-55885701;chr7:55885641-5588576 1;chr7:55885701-55885821;chr7:55885761-55885881;chr7:55885821-55885941;chr7:55885881-55886001;chr7:55885941-55886061;ch r7:55887081-55887201;chr7:55887141-55887261;chr7:55887201-55887321;chr7:55887261-55887381;chr7:55887321-55887441;chr7:5 5887381-55887501;chr7:55887441-55887561;chr7:55887501-55887621;chr9:69037474-69037594;chr9:69037534-69037654;chr9:690375 94-69037714;chr9:69037894-69038014;chr9:69037954-69038074;chr9:69038014-69038134;chr9:69038074-69038194;chr9:69038134-69038254 ;chr9:69038194-69038314;chr9:69038254-69038374;chr9:69038314-69038434;chr9:69038374-69038494;chr9:69038434-69038554;chr9:690387 34-69038854;chr9:69038794-69038914;chr9:69038854-69038974;chr9:69039214-69039334;chr9:69039274-69039394;chr9:130679945-13068006 5;chr9:130680665-130680785;chr9:130680725-130680845;chr9:130680785-130680905;chr9:130680845-130680965;chr9:130680905-130681025;chr9:130680965-130681085; chr9:130681025-130681145; chr9:130681085-130 681205;chr9:130681145-130681265;chr9:130681205-130681325;chr9:1306812 65-130681385; chr9:130681325-130681445; chr9:130681385-130681505; chr9:1 30681445-130681565; chr9:130681505-130681625; chr9:130681565-130681685;

[0032] 3) The 3× tiling probe design region includes the following gene loci:

[0033] STR gene loci: ATXN3, STARD7, DIP2B;

[0034] VNTR gene locus: PLIN4;

[0035] The specific sequences of 3× tiling probes are (a total of 109):

[0036] chr12:50502921-50503041;chr12:50502961-50503081;chr12:50503001-50503121;chr12:50503041-50503161;chr12:50503081-50503201;chr12:50503761-50503881;chr12:50503801-50503921;chr12:50503841-50503961;chr12:50503881-50504001;chr12:50504521-50504641;chr12:50504561-50504681;chr12:50504601-50504721;chr12:50504641-50504761;chr12:50504681-50504801;chr12:50504721-50504841;chr12:50504761-50504881;chr12:50504801-50504921;chr12:50504841-50504961;chr12:50504881-50505001;chr12:50504921-50505041;chr12:50504961-50505081;chr14:92068930-92069050;chr14:92068970-92069090;chr14:92069010-92069130;chr14:92070050-92070170;chr14:92070090-92070210;chr14:92070130-92070250;chr14:92070170-92070290;chr14:92070210-92070330;chr14:92070570-92070690;chr14:92070610-92070730;chr14:92070730-92070850;chr14:92070770-92070890;chr14:92070810-92070930;chr14:92070850-92070970;chr14:92070890 -92071010;chr14:92070930-92071050;chr14:92070970-92071090;chr19:4508527-4508647;chr19:4508567-4508687;chr19:4508607-4508727;chr19:4508647-4508767;<h2 style=";text-align:left;direction:ltr">chr19:4508687-4508807;chr19:4508727-4508847;chr19:4508767-4508887;chr19:4508807-4508927;chr19:4508847-4508967;chr19:450888 7-4509007;chr19:4508927-4509047;chr19:4508967-4509087;chr19:4510327-4510447;chr19:4510367-4510487;chr19:4510407-4510527;chr 19:4510447-4510567;chr19:4513501-4513621;chr19:4513541-4513661;chr19:4513581-4513701;chr19:4513621-4513741;chr19:4513661-45 13781;chr19:4513701-4513821;chr19:4513741-4513861;chr19:4513781-4513901;chr19:4513821-4513941;chr19:4513861-4513981;chr19:4 513901-4514021;chr19:4513941-4514061;chr19:4513981-4514101;chr19:4514021-4514141;chr19:4514061-4514181;chr19:4514101-451422 1;chr19:4515541-4515661;chr2:96194975-96195095;chr2:96195015-96195135;chr2:96195055-96195175;chr2:96195095-96195215;chr2:96 195135-96195255;chr2:96195175-96195295;chr2:96195215-96195335 ;chr2:96195255-96195375;chr2:96195295-96195415;chr2:96195335-9 6195455;chr2:96195375-96195495;chr2:96195415-96195535;chr2:96 195455-96195575;chr2:96195495-96195615;chr2:96195535-96195655;chr2:96195575-96195695; chr2:96195615-96195735; chr2:96195655-961957 75; chr2:96195695-96195815; chr2:96195735-96195855; chr2:96197415-961 97535;chr2:96197455-96197575;chr2:96197495-96197615;chr2:96197535- 96197655;chr2:96197575-96197695;chr2:96197935-96198055;chr2:961979 75-96198095; chr2:96198375-96198495; chr2:96198415-96198535; chr2:961 98455-96198575;chr2:96198495-96198615;chr2:96198775-96198895;chr2: 96198815-96198935;chr2:96198855-96198975;chr2:96198895-96199015;ch r2:96198935-96199055; chr2:96199055-96199175; chr2:96199095-96199215;

[0037] 4) The 4× tiling probe design region includes the following gene loci:

[0038] STR gene loci: ATXN10, ZNF713;

[0039] The specific sequences of the 4× tiling probes are (37 in total):

[0040] chr22:45795329-45795449; chr22:45796769-45796889; chr22:45796799-45796919; chr22:45796829-45796949; chr22:457968 59-45796979;chr22:45796889-45797009;chr22:45796919-45797039;chr22:45796949-45797069;chr22:45796979-45797099;c hr22:45797009-45797129; chr22:45797039-45797159; chr22:45797069-45797189; chr22:45797099-45797219; chr22:4579712 9-45797249;chr22:45797159-45797279;chr22:45797189-45797309;chr22:45797219-45797339;chr7:55887874-55887994;chr 7:55887904-55888024;chr7:55887934-55888054;chr7:55887964-55888084;chr7:55887994-55888114;chr7:55888024-55888 144;chr7:55888054-55888174;chr7:55888084-55888204;chr7:55888114-55888234;chr7:55888204-55888324;chr7:55888234 -55888354;chr7:55888264-55888384;chr7:55888294-55888414;chr7:55889434-55889554;chr7:55889464-55889584;chr7:5 5889494-55889614; chr7:55889524-55889644; chr7:55889554-55889674; chr7:55889584-55889704; chr7:55889614-55889734.

[0041] The probes of the present invention are designed based on the human reference genome version hg38, wherein each probe in the probe set is at least one of a double-stranded DNA, a single-stranded DNA, and a single-stranded RNA; both the forward sequence and the reverse complementary sequence of the double-stranded DNA probe can be used to capture dynamically mutated STR and VNTR gene sites.

[0042] Each probe in the probe set of the present invention is labeled for signal amplification and luminescence detection after the probe specifically binds to fragmented DNA, and includes at least one of biotin, avidin, avidin, an antibody, or a chemical coupling agent, preferably biotin.

[0043] In a second aspect, the present invention provides a method for constructing a single-molecule long-read sequencing library, comprising the following steps:

[0044] 1) Randomly fragment the DNA;

[0045] 2) Perform end repair on the fragmented DNA and ligate with pre-library adapters to obtain a pre-library;

[0046] 3) hybridizing the prelibrary with the probe set of the present invention in the absence of a blocking sequence to obtain a hybridization product;

[0047] 4) Using magnetic beads to capture the hybridization products and after washing, PCR amplification is performed to obtain a capture library;

[0048] 5) Construction of third-generation sequencing library.

[0049] In one embodiment, genomic DNA can come from a variety of sources, such as blood, blood spots, semen, semen stains, bones, hair, saliva, saliva stains, sweat, tissue, and the like. Those skilled in the art are aware of methods for fragmenting genomic DNA, such as by sonication, mechanical fragmentation, or enzymatic digestion, and the fragmentation conditions can be adjusted according to actual needs.

[0050] In one embodiment, the fragmented DNA is 5-15 kb in length, preferably 7-10 kb in length.

[0051] In one embodiment, the pre-library adapter has a Y-shaped structure and can be an overhanging T-adapter or a blunt-end adapter. A "Y-adapter" refers to a adapter formed by annealing two incompletely complementary DNA single strands. One end of the adapter forms a double strand due to base complementarity between the two strands, while the other end does not form a double strand due to incomplete base complementarity between the two strands. For example, the sequences of the two strands of the Y-adapter that can be used in the present invention are as follows:

[0052] F:

[0053] 5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATC-3'(SEQ ID NO.1)

[0054] R: (5' end p represents phosphorylation modification)

[0055] 5'-pGATCGGAAGAGCACACGTCTGAACTCCAGTCACATCTCGTATGCCGTCTTCTGCTTG-3'(SEQ ID NO.2)

[0056] The underlines indicate the base-pairing parts of the two single strands.

[0057] In the context of the present invention, a "blocking sequence" refers to a sequence used to block an adapter and / or tag sequence, including sequences designed to be reverse complementary to the adapter and / or tag sequence. In some embodiments, to increase the blocking effect of the blocking sequence, special modifications are added to the ends of the blocking sequence, such as reverse dT modification, amino modification, ddNTP modification (including ddCTP, ddATP, ddGTP and ddTTP), spacer modification, hypoxanthine modification, random base modification, etc.

[0058] Based on the findings of the present inventors, in the absence of pre-capture PCR amplification and the use of a Y-type connector, good capture efficiency can be achieved without the need to add any blocking sequence to the hybridization system. Therefore, in one embodiment, a pre-library can be obtained without the need for pre-capture PCR amplification. In another embodiment, the system for hybridization includes a hybridization buffer, an enhancement solution, human Cot-1 DNA, and a hybridization probe, but does not include a blocking sequence. Those skilled in the art can adjust the conditions for hybridization capture, such as hybridization temperature, hybridization time, hot wash temperature, and number of hot washes, according to actual needs. The general principles of designing and preparing hybridization probes are also well known to those skilled in the art.

[0059] In one embodiment, the magnetic beads are used to capture hybridization products, preferably streptavidin magnetic beads.

[0060] In one embodiment, the post-capture PCR amplification system includes an amplification enzyme, an amplification buffer, a PCR additive, amplification primers, and the post-magnetic bead capture product. The amplification enzyme has high fidelity and excellent amplification properties, making it suitable for amplifying complex templates such as long fragments and those with high GC / AT content. The PCR additive can reduce secondary structure, enhance enzyme stability, or inhibit nonspecific amplification, and is selected from at least one of dimethyl sulfoxide, betaine, bovine serum albumin, and formamide. The amplification primers have a final concentration range of 0.1-1 μM, are phosphorylated, and are compatible with pre-library adapters. The amplification program includes: 94°C for 2-4 minutes; 98°C for 10-15 seconds, Tm for 30 seconds, 68°C for 5-10 minutes, 15-25 cycles; and 68°C for 5-10 minutes.

[0061] In one embodiment, the post-capture PCR amplification product and the major band of the third-generation sequencing library are approximately 8 kb.

[0062] In a specific embodiment, SMRT sequencing is performed using multiple dynamic mutation sequencing libraries. The data from a single chip indicate that the overall insert length is concentrated between 5 and 10 kb, as shown in FIG1 .

[0063] In a third aspect, the present invention provides a device for detecting the number of repeats of STR and VNTR gene loci, comprising:

[0064] 1) Capture module: used to construct capture library;

[0065] 2) Library construction module: used to construct single-molecule long-read sequencing libraries;

[0066] 3) Sequencing and analysis module: Perform third-generation sequencing on the sequencing library and perform data analysis to obtain sequence information of the target repetitive region.

[0067] In one embodiment, the probe set used in the apparatus of the present invention to construct a capture library is selected from the probe sets described above.

[0068] In one embodiment, the method for constructing a capture library and a single-molecule long-read sequencing library using the apparatus of the present invention is selected from the sequencing library construction method described above.

[0069] In a specific embodiment, the single-molecule long-read sequencing platform is selected from the Pacific Biosciences SMRT sequencing platform based on single-molecule real-time sequencing technology and the Oxford Nanopore nanopore single-molecule sequencing platform.

[0070] In one embodiment, the data analysis process of the device of the present invention includes:

[0071] 1) After the third-generation sequencing data is split into samples, it is aligned with the human genome reference sequence;

[0072] 2) retain the uniquely aligned sequences and screen the target regions;

[0073] 3) Count the number of target repeat region sequences, capture efficiency and coverage depth.

[0074] In one embodiment, the method for detecting the number of repetitions of the STR gene locus is:

[0075] 1) Based on the sequence of the target region, analyze the repeat region, calculate the number of repeats and supporting sequences, and determine pathogenicity based on the information included in the database;

[0076] 2) Based on the specific sequence alignment of the 5' and 3' flanking regions of the target repeat region, the repeat region sequence is cut, the number of repeat units and typing clustering are analyzed, the repeat structure of each allele is obtained, and the number of repeats for each allele and the number of sequences supporting the allele are counted.

[0077] In a specific embodiment, if the two alleles of the STR gene locus detected by the above method have the same number of repeats, the haplotypes are split according to the SNVs of the flanking regions of the repeat region; for the two alleles with the same repeat structure, the number of repeats and the number of supporting sequences are counted separately.

[0078] In a specific embodiment, if the STR gene locus detected by the above method is positive for high repeat interruption, the unilateral repeat sequence is aligned with the flanks of the repeat region, the upstream or downstream of the repeat region is located, sheared and filtered, the number of repeat units is calculated, and the pathogenicity determination is supplemented.

[0079] In a specific embodiment, in the method for detecting the number of repetitions of STR gene loci, the output value of the number of allele repetitions can be selected from the following statistics: mean, third quartile, median, mode, or peak value returned by kernel density estimation method, preferably median.

[0080] In one embodiment, the method for detecting the number of repetitions of the VNTR gene locus is:

[0081] 1) Due to the large number of polymorphisms in VNTR repeat units, the detection of repeat regions in VNTR gene loci requires the construction of a repeat type list for each VNTR gene based on the existing sample set;

[0082] 2) Based on the specific sequence alignment of the 5' and 3' flanking regions of the target repeat region, the repeat region sequence is cut, and the repeat units are identified from the built-in VNTR gene repeat type list. Polymorphisms of the same type are combined to calculate the number of repeats;

[0083] 3) Similar to STR repeats, typing and clustering are performed to count the number of repeats and supporting sequences for each allele.

[0084] In a specific embodiment, in the method for detecting the number of repetitions of the VNTR gene loci, the output value of the number of allele repetitions can be selected from the following statistics: mean, third quartile, median, mode or peak value returned by the kernel density estimation method, preferably the median.

[0085] The dynamic mutation STR and VNTR gene locus repeat count detection method described in this invention not only reports the repeat count of known pathogenic repeat units, but also simultaneously reports the repeat counts of other repeat units within the repeat region. These other repeat units are derived from a comprehensive search of databases and published articles. This method can reveal the relationship between repeat units that have been reported but not yet included in databases or whose pathogenicity is not yet clearly established and the disease, thereby facilitating the comprehensive exploration of repeat mutations that may cause diseases.

[0086] In a fourth aspect, the present invention provides a method for detecting the number of repeats of dynamically mutated STR and VNTR gene loci, comprising the following steps:

[0087] 1) Perform random fragmentation, end repair, and pre-library adapter ligation on the DNA sample to obtain a pre-library;

[0088] 2) In the absence of a blocking sequence, the probe set is used to hybridize with the prelibrary, and the hybridization products are captured using magnetic beads, and then PCR amplified after washing to obtain a captured library;

[0089] 3) Construct a third-generation sequencing library, perform third-generation sequencing and analyze the sequencing data to obtain target sequence information.

[0090] In one embodiment, the method of the present invention for constructing a pre-library, a capture library, and a third-generation sequencing library is selected from the method for constructing a single-molecule long-read sequencing library as described above.

[0091] In one embodiment, the method for analyzing sequencing data according to the present invention is selected from the data analysis methods described above.

[0092] In a fifth aspect, the present invention provides a kit for detecting dynamically mutated STR and VNTR gene loci, comprising the following components:

[0093] 1) The probe set of the present invention;

[0094] 2) Reagents for constructing prelibrary;

[0095] 3) Reagents used for hybridization capture, excluding blocking sequences;

[0096] 4) Reagents for PCR amplification;

[0097] 5) Reagents for constructing third-generation sequencing libraries.

[0098] In one embodiment, the reagents for constructing the prelibrary include a repair enzyme, a repair buffer, a ligase, a ligation buffer, and a prelibrary adapter. In a specific embodiment, the prelibrary adapter is a Y-shaped structure, which can be an overhanging T adapter or a blunt-end adapter.

[0099] In one embodiment, the hybridization capture reagent includes hybridization buffer, enhancement solution, human Cot-1 DNA, the probe set of the present invention, magnetic beads and washing reagent, but does not include a blocking sequence.

[0100] In one embodiment, the magnetic beads are used to capture hybridization products, preferably streptavidin magnetic beads.

[0101] In one embodiment, the PCR amplification reagents include an amplification enzyme, an amplification buffer, a PCR additive, and amplification primers. The amplification enzyme has high fidelity and excellent amplification properties, making it suitable for amplifying complex templates such as long fragments and those with high GC / AT content. The PCR additive can reduce secondary structure, enhance enzyme stability, or inhibit nonspecific amplification, and is selected from at least one of dimethyl sulfoxide, betaine, bovine serum albumin, and formamide. The amplification primers have a final concentration range of 0.1-1 μM, are phosphorylated, and are compatible with pre-library adapters.

[0102] In one embodiment, the reagents for constructing a third-generation sequencing library include a repair enzyme, a ligase, an exonuclease and a matching buffer, as well as a third-generation sequencing adapter.

[0103] In a sixth aspect, the present invention also provides the use of the probe group or the single-molecule long-read sequencing library construction method or the STR and VNTR gene locus repetition number detection device or the STR and VNTR gene locus repetition number detection method in the preparation of a kit for detecting dynamically mutated STR and VNTR gene loci.

[0104] In one embodiment, the STR and VNTR gene loci kit is suitable for detecting one or more of the following gene loci:

[0105] 1) STR loci (69): ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7, ATXN8OS / ATXN8, ATXN10, PPP2R2B, TBP, BEAN1 / TK2, NOP56, DAB1, ATN1, FXN, GLS, RFC1, AR, DMPK, CNBP, LRP12, GIPC1, NUTM2B-AS1 / LOC642361, PABPN1, C9orf72, NIPA1, TAF1, DMD, VW A1, SPAST, CSTB, SAMD12, STARD7, MARCHF6, YEATS2, TNRC6A, RAPGEF2, NOTCH2NLC, FMR1, AFF2, AFF3, ZNF713, C11ORF80, DIP2B, CBL, HTT, JPH3, PRDM12, PRNP, SOX3, ARX, ZIC2, HOXD13, EIF4A3, RUNX2, HOXA13, FOXL2, ZIC3, TBX1, XYLT1, COMP, TCF4, PHOX2B, FGF14, RILPL1, THAP11, POLG;

[0106] 2) VNTR gene loci (4): CACNA1C, PLIN4, ABCA7, and WDR7.

[0107] In one embodiment, the probe group of the present invention or the detection device for the number of repetitions of the STR and VNTR gene loci or the detection kit for the dynamic mutation STR and VNTR gene loci can be used to detect one or more of the following diseases: spinocerebellar ataxia type 1 (SCA1), spinocerebellar ataxia type 2 (SCA2), spinocerebellar ataxia type 3 (SCA3), spinocerebellar ataxia type 6 (SCA6), spinocerebellar ataxia type 7 (SCA7), spinocerebellar ataxia type 8 (SCA8), spinocerebellar ataxia type 12 (SCA12), spinocerebellar ataxia type 17 (SCA17), spinocerebellar ataxia type 36 (SCA36), Dentatorubropallidular atrophy with Lewy bodies (DRPLA), Kennedy disease (SBMA), myotonic dystrophy type 1 (DM1), Huntington disease (HD), Huntington-like disease 2 (HDL2), oculopharyngeal muscular dystrophy (OPMD), spinocerebellar ataxia type 37 (SCA37), Friedreich's ataxia (FRDA), generalized developmental delay, progressive ataxia, elevated glutamine (GDPAG), fragile X-associated tremor / ataxia syndrome (FXTAS), fragile X syndrome (FXS), fragile X-associated ovarian insufficiency (FXPOI), fragile X-associated psychiatric disorder (FXAND), fragile XE syndrome (FRAXE), KINSSHIP Syndrome (FRA2A), mental retardation FRA11A type (FRA11A), mental retardation FRA12A type (FRA12A), Jacobsen syndrome (FRA11B), familial adult myoclonic epilepsy type 1 (FAME1), familial adult myoclonic epilepsy type 6 (FAME6), familial adult myoclonic epilepsy type 7 (FAME7), early infantile epileptic encephalopathy type 1 (ELEE1) / Partington mental retardation syndrome (PRTS) / X-linked intellectual disability with or without seizures (MRXARX), myotonic dystrophy type 2 (DM2), neuronal intranuclear inclusion disease (NIID), hereditary essential tremor type 6 (ETM6), oculopharyngeal myopathy 3 Type (OPDM3), oculopharyngeal myopathy type 1 (OPDM1), oculopharyngeal myopathy type 2 (OPDM2), oculopharyngeal leukoencephalopathy with leukoencephalopathy (OPML1), amyotrophic lateral sclerosis 1 (ALS1), Duchenne muscular dystrophy (DMD), hereditary motor neuropathy (HMN), hereditary sensory and autonomic neuropathy type 8 (HSAN8), spastic paraplegia 4 (SPG4), Richieri-Costa-Pereira syndrome (RCPS), Creutzfeldt-Jakob disease (CJD), Fuchs endothelial corneal dystrophy type 3 (FECD3), polydactyly type 1, brachydactyly and cleidocranial dysplasia (CCD), hand-foot-genital syndrome (HFGS), blepharophimosis microsomia syndrome (BPES),Holoprosencephaly type 5 (HPE5), monogenic long hormone deficiency with mental retardation (MRGH), X-linked hypopituitarism (PHPX), congenital central hypoventilation syndrome (CCHS), combined congenital malformation (VACTERL), oculo-auriculo-spinal malformation (OVAS), tetralogy of Fallot (ToF), epiphyseal dysplasia type 1 (EDM1), pseudoachondroplasia (PSACH), oculopharyngeal myopathy type 4 (OPDM4), spinocerebellar ataxia tarda-27B (SCA27B), Baratela-Scott syndrome (BSS), Alzheimer's disease (AD), schizophrenia and bipolar disorder (BD / SCZ), myopathy with ubiquitin-positive autophagy-rimmed vacuoles (MRUPAV) ), X-linked dystonia-Parkinson's disease (XDP), amyotrophic lateral sclerosis 3 (ALS3), spinocerebellar ataxia type 10 (SCA10), spinocerebellar ataxia type 31 (SCA31), cerebellar ataxia with neuropathy and vestibular areflexia syndrome (CANVAS), autism spectrum disorder (FRA7A), progressive myoclonic epilepsy type 1 (EPM1), familial adult myoclonic epilepsy type 2 (FAME2), familial adult myoclonic epilepsy type 3 (FAME3), familial adult myoclonic epilepsy type 4 (FAME4), frontotemporal dementia / amyotrophic lateral sclerosis (C9orf72-FTD / ALS), spinocerebellar ataxia, Friedreich's ataxia (FRDA).

[0108] The present invention discloses a probe set, detection equipment and kit for detecting dynamically mutated STR and VNTR gene loci. Based on targeted capture of long fragments and long-read sequencing technology, the present invention can detect the target repeat region sequence information of all targeted gene loci in a single sample at one time, overcoming the shortcomings of traditional detection technology of single disease single detection, no target repeat region sequence information, limited detectable range and long time required for differential diagnosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0109] Figure 1: Sequencing library data.

[0110] Figure 2: Flowchart of sequencing library construction.

[0111] Figure 3: Sequencing library fragment analysis. Lower refers to the Lower Marker.

[0112] Figure 4: Target repeat region information of the PLIN4 gene in sample 31.

[0113] Figure 5: Target repeat region information of the PLIN4 gene in sample 32.

[0114] Figure 6: Target repeat region information of the PLIN4 gene in sample 33. DETAILED DESCRIPTION

[0115] The invention generally described herein will be more readily understood by reference to the following examples, and those skilled in the art may refer to the contents herein and appropriately modify the process parameters for implementation. It is particularly important to note that all similar substitutions and modifications will be apparent to those skilled in the art and are considered to be within the scope of protection of the present invention. The following examples are provided by way of illustration and are not intended to limit the present invention. These examples are not intended to represent that the experiments below are all or only the experiments performed.

[0116] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials, reagents, etc. used are all available from commercial sources unless otherwise specified.

[0117] The present invention provides probe sets, detection devices, and kits for detecting dynamically mutated STR and VNTR loci. By leveraging targeted capture of long fragments and long-read sequencing technology, this technology enables simultaneous detection of 73 STR and VNTR loci in a single sample, encompassing 79 diseases. This facilitates rapid, accurate, and comprehensive molecular diagnosis in clinics. The correspondence between the 73 dynamically mutated loci and the 79 diseases involved is shown in Table 1.

[0118] Table 1: Correspondence between 73 dynamic mutation loci and 79 diseases involved

[0119] Example 1: Construction of a third-generation sequencing library and testing the effect of different DNA starting amounts on the main band of the library

[0120] The third-generation sequencing library construction process is shown in Figure 2, and the specific construction method is as follows:

[0121] 1. Genomic DNA Fragmentation

[0122] Genomic DNA was fragmented using g-TUBE (Covaris, Cat. No. 520079). Starting amounts of 2 μg, 3 μg, and 4 μg of DNA from the same sample were used. After fragmentation, the fragmented DNA was purified using AMPure PB magnetic beads.

[0123] 2. End repair

[0124] End repair was performed using the end repair reagent (Enzymatics, Cat. No. Y9140-HC-L) in the reaction system shown in Table 2. The reaction procedure was as follows: 25°C for 30 minutes, 75°C for 20 minutes, and then maintained at 4°C.

[0125] Table 2: End-repair systems

[0126] 3. Prelibrary adapter ligation

[0127] Use the ligase reagent (Enzymatics, Cat. No. L6030-HC-L) to prepare the reaction system shown in Table 3 and complete the ligation reaction. The reaction schedule was: 20°C for 30 minutes, followed by a hold at 4°C. After the ligation reaction, AMPure PB magnetic beads were used for purification.

[0128] Table 3: Connection system

[0129] 4. Pre-library hybridization

[0130] Add the hybridization reagent (IDT, Catalog No. 1072281) listed in Table 4 to the purified product, mix thoroughly, and incubate at room temperature for 10 minutes. After incubation, transfer the supernatant to a new low-adhesion 0.2 mL centrifuge tube and add 4 μL of the dynamic mutation probe set provided by the present invention. Mix thoroughly, centrifuge briefly, and run the following cycle on a PCR instrument: 95°C for 5 minutes; 47°C for 17 hours; then hold at 47°C.

[0131] Table 4: Hybridization system

[0132] 5. Magnetic Bead Capture and Washing

[0133] The hybridization products were captured using streptavidin magnetic beads and washed with heat and room temperature (IDT, Cat. No. 1072281). After washing, the beads were resuspended in enzyme-free water.

[0134] 6. PCR Amplification

[0135] Add the reaction system shown in Table 5 (TOYOBO, Catalog No. KFX-201) to the above product, mix thoroughly, centrifuge briefly, and run the program shown in Table 6. After the PCR reaction, purify using AMPure PB magnetic beads.

[0136] Table 5: PCR reaction system

[0137] Table 6: PCR reaction program

[0138] 7. Construction of third-generation sequencing library

[0139] A third-generation sequencing library was constructed using a kit (PacBio, catalog number 102-088-900). The reagents included repair enzyme, ligase, exonuclease, and matching buffer, as well as third-generation sequencing adapters, to ultimately obtain a sequencing library.

[0140] 8. Library QC and Results Presentation

[0141] The library was analyzed using the Agilent 4200 TapeStation system and genomic DNA reagents (Agilent, 5067-5365, 5067-5366). The results showed that libraries could be constructed with an input amount of 2-4 μg DNA, and the major bands of the libraries were approximately 8 kb, as shown in Table 7 and Figure 3.

[0142] Table 7: Library fragment analysis results

[0143] Example 2: Construction of a third-generation sequencing library to verify STR gene locus detection performance

[0144] Referring to the library construction method of Example 1, sequencing libraries were constructed using 27 clinical samples and 3 Coriell human genome reference samples. The median repeat counts of different repeat units were calculated and compared with the results of RP-PCR or dmWES detection or Coriell reference results. The results are shown in Table 8. The median repeat counts detected by the present method are consistent with those of RP-PCR, dmWES detection, and Coriell reference results.

[0145] The method of the present invention reports not only the repeat counts of known pathogenic repeat units, but also the repeat counts of other repeat units within the repeat region. These other repeat units, derived from comprehensive searches of databases and published articles, can reveal the relationship between repeat units that have been reported but not yet included in databases or whose pathogenicity is not yet clearly established and the disease, thereby facilitating a comprehensive exploration of repeat mutations that may cause disease.

[0146] Table 8: STR gene locus detection results. In Sample 1 tested using the present method, "ATXN1|CAG,CAT 27_1;60_1" indicates that one autosomal ATXN1 allele has 27 CAG units and 1 CAT unit, while the other allele has 60 CAG units and 1 CAT unit. The number of replicates detected by RP-PCR may include other interrupted sequences. For example, in Sample 1, "ATXN1|CAG,CAT 28;61" indicates that one autosomal ATXN1 allele has 28 CAG units (including CAT units) and the other allele has 61 CAG units (including CAT units). The same applies below. " / " indicates that the allele was not tested. Note: When the detection limit of RP-PCR or dmWES is not exceeded, the results are considered consistent if the number of repetitions of the repeat unit is within ±2. When the detection limit is exceeded, the theoretical pathogenicity determination is consistent. Samples 28-30 are Coriell standard samples NA07537 (FMR1|CGG 28-29; >200), NA07862 (FMR1|CGG 501-550), and NA09237 (FMR1|CGG931-940), respectively. The results are considered consistent if the theoretical pathogenicity determination is consistent.

[0147] Example 3: Construction of a third-generation sequencing library to verify VNTR gene locus detection performance

[0148] Referring to the library construction method described in Example 1, sequencing libraries were constructed using clinical samples from three hospitals with known long-range PCR results. The target repeat region of the VNTR gene locus PLIN4 was detected and compared with the long-range PCR amplification results. The results, shown in Table 9 and Figures 4-6, show that the repeat region length detected by the present method is consistent with the long-range PCR amplification results. Samples 32 and 33 both showed bands indicating abnormal repeat amplification.

[0149] Table 9: PLIN4 gene locus detection results Note: The results are considered consistent when the target repeat region length is close to the PCR product size.

Claims

1. A probe set for detecting dynamically mutated STR and VNTR gene loci, targeting the following loci: 1) STR gene loci (69): ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7, ATXN8OS / ATXN8, ATXN10, PPP2R2B, TBP, BEAN1 / TK2, NOP56, DAB1, ATN1, FXN, GLS, RFC1, AR, DMPK, CNBP, LRP12, GIPC1, NUTM2B-AS1 / LOC642361, PABPN1, C9orf72, NIPA1, TAF1, DMD, VWA1, SPAST, CSTB, SAMD1 2. STARD7, MARCHF6, YEATS2, TNRC6A, RAPGEF2, NOTCH2NLC, FMR1, AFF2, AFF3, ZNF713, C11ORF80, DIP2B, CBL, HTT, JPH3, PRDM12, PR NP, SOX3, ARX, ZIC2, HOXD13, EIF4A3, RUNX2, HOXA13, FOXL2, ZIC3, TBX1, XYLT1, COMP, TCF4, PHOX2B, FGF14, RILPL1, THAP11, POLG; 2) VNTR gene loci (4): CACNA1C, PLIN4, ABCA7, and WDR7.

2. The probe set according to claim 0, which meets the following requirements: 1) Covering the 5' flanking region and the 3' flanking region of the STR and VNTR gene loci of claim 0, the main body adopts a 1× tiling coverage method to design probes; for areas that are not suitable for designing probes, such as those with poor sequence specificity, no probes are designed; when the probe covers a small area, the probe density is increased according to the following principles: If the bases covered by the probes of the 2kb regions on both sides are between 700-1000bp, the probes are designed by 2× tiling coverage; if the bases covered by the probes of the 2kb regions on both sides are between 500-700bp, the probes are designed by 3× tiling coverage; if the bases covered by the probes of the 2kb regions on both sides are less than 500bp, the probes are designed by 4× tiling coverage; 2) The probe length is 80–120 bp, and the GC content is preferably 40%-60%.

3. According to claim 0 or 2, for the STR gene locus, when the probe is designed by 4× tiling coverage, if the target repeat region of the gene locus can be used to design a probe, it is preferred to design the probe by 1× tiling coverage; or design the probe by 1× tiling coverage in the 2nd-3kb region of the 5' flanking region and the 3' flanking region of the gene locus.

4. According to the probe set of claim 0 or 2, for the VNTR gene locus, if the target repeat region of the gene locus can be designed with a probe, the probe is preferably designed by a 1× tiling coverage method; or the probe is designed by a 1× tiling coverage method in the 2nd-3kb region of the 5' flanking region and the 3' flanking region of the gene locus.

5. The probe set according to any one of claims 0 to 4, comprising all or at least one of the following 2230 sequences: 1) The 1× tiling probe design region includes the following gene loci: STR loci: ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7, ATXN8OS / ATXN8, ATXN10, PPP2R2B, TBP, BEAN1 / TK2, NOP56, DAB1, ATN1, FXN, GLS, RFC1, AR, DMPK, CNBP, LRP12, GIPC1, PABPN1, C9orf72, NIPA1, TAF1, DMD, VWA1, CSTB, SAMD12, MARCHF6, YEA TS2, TNRC6A, RAPGEF2, NOTCH2NLC, FMR1, AFF2, AFF3, ZNF713, C11ORF80, DIP2B, CBL, HTT, JPH3, PRDM12, PRNP, SOX3, AR X, ZIC2, HOXD13, EIF4A3, RUNX2, HOXA13, FOXL2, ZIC3, TBX1, XYLT1, COMP, TCF4, PHOX2B, FGF14, RILPL1, THAP11, POLG; VNTR gene loci: CACNA1C, PLIN4, ABCA7, WDR7; 1× tiling probe specific probe sequence (a total of 1867): chr1:1433798-1433918; chr1:1433915-1434035; chr1:1434033-1434153; chr1:1434150-1434270; chr1:1434271-14 34391;chr1:1434388-1434508;chr1:1434505-1434625;chr1:1434622-1434742;chr1:1434743-1434863;chr1:1434 860-1434980;chr1:1434978-1435098;chr1:1435095-1435215;chr1:1435216-1435336;chr1:1435333-1435453;chr 1:1435451-1435571;chr1:1435568-1435688;chr1:1435689-1435809;chr1:1435806-1435926;chr1:1435924-14360 44;chr1:1436041-1436161;chr1:1436162-1436282;chr1:1436279-1436399;chr1:1436396-1436516;chr1:1436513 -1436633;chr1:1436634-1436754;chr1:1436751-1436871;chr1:1436869-1436989;chr1:1436986-1437106;chr1:1 437107-1437227;chr1:1437224-1437344;chr1:1437342-1437462;chr1:1437459-1437579;chr1:1437580-1437700; chr1:1437698-1437818; chr1:57365392-57365512; chr1:57365513-57365633; chr1:57365629-57365749; chr1:5736 5746-57365866;chr1:57365862-57365982;chr1:57365983-57366103;chr1:57366099-57366219;chr1:57366216-57366336;chr1:57366332-57366452;chr1:57366453-57366573;chr1:57366569-57366689;chr1:57366686-57366806;chr1:57366802-57366922;chr1:57367241-57367361;chr1:57367358-57367478;chr1:57367474-57367594;chr1:57367595-57367715;chr1:57367711-57367831;chr1:57367828-57367948;chr1:57367944-57368064;chr1:57368065-57368185;chr1:57368181-57368301;chr1:57368768-57368888;chr1:57368884-57369004;chr1:57369005-57369125;chr1:149390842-149390962;chr1:149391545-149391665;chr1:149391661-149391781;chr1:149391782-149391902;chr1:149391898-149392018;chr1:149392015-149392135;chr1:149392131-149392251;chr1:149392252-149392372;chr1:149392368-149392488;chr1:149392485-149392605;chr1:149392601-149392721;chr1:149392722-149392842;chr11:66742937-66743057;chr11:66743055-66743175;chr11:66743172-66743292;chr11:66743293-66743413;chr11:66743411-66743531;chr11:66743528-66743648;chr11:66743646-66743766;chr11:66743767-66743887;chr11:66744119-66744239;chr11:66744240-66744360;chr11:66744357-66744477;chr11:66744475-66744595;chr11:66744592-66744712;chr11:66744713-66744833;chr11:66744831-66744951;chr11:66744948-66745068;chr11:66745066-66745186;chr11:66745187-66745307;chr11:66745304-66745424;chr11:66745422-66745542;chr11:66745539-66745659;chr11:66745660-66745780;chr11:66746133-66746253;chr11:119204643-119204763;chr11:119204764-119204884;chr11:119204882-119205002;chr11:119205000-119205120;chr11:119205118-119205238;chr11:119205239-119205359;chr11:119205357-119205477;chr11:119205474-119205594;chr11:119205592-119205712;chr11:119205713-119205833;chr11:119205831-119205951;chr11:119205948-119206068;chr11:119206066-119206186;chr11:119206187-119206307;chr11:119206305-119206425;chr11:119206423-119206543;chr11:119206541-119206661;chr11:119206662-119206782;chr11:119206780-119206900;chr11:119206897-119207017;chr11:119207015-119207135;chr11:119207136-11920725; 6;chr11:119207254-119207374;chr11:119207371-119207491;chr11:119207489-119207609;chr11:119207610-119207730;chr11:119207728-119207848;chr11:119207845-119207965;chr11:119207963-119208083;chr11:119208084-119208204;chr11:119208203-119208323;chr12:2253791-2253911;chr12:2253907-2254027;chr12:2254024-2254144;chr12:2254140-2254260;chr12:2254261-2254381;chr12:2254377-2254497;chr12:2254494-2254614;chr12:2254610-2254730;chr12:2254731-2254851;chr12:2254847-2254967;chr12:2254964-2255084;chr12:2255080-2255200;chr12:2255201-2255321;chr12:2255317-2255437;chr12:2255434-2255554;chr12:2255550-2255670;chr12:2255671-2255791;chr12:2256090-2256210;chr12:2256206-2256326;chr12:2256323-2256443;chr12:2256439-2256559;chr12:2256560-2256680;chr12:2256676-2256796;chr12:2256793-2256913;chr12:2256909-2257029;chr12:2257030-2257150;chr12:2257146-2257266;chr12:2257263-2257383;chr12:2257616-2257736;chr12:2257733-2257853;chr12:2257849-2257969;chr12:2257970-2258090;chr12:2255791-2255911;chr12:2255880-2256000;chr12:2255970-2256090;chr12:6934716-6934836;chr12:6935193-6935313;chr12:6935313-6935433;chr12:6935432-6935552;chr12:6935552-6935672;chr12:6935671-6935791;chr12:6935790-6935910;chr12:6935910-6936030;chr12:6936029-6936149;chr12:6936148-6936268;chr12:6936268-6936388;chr12:6936387-6936507;chr12:6936506-6936626;chr12:6936626-6936746;chr12:6936745-6936865;chr12:6936865-6936985;chr12:6936984-6937104;chr12:6937103-6937223;chr12:6937223-6937343;chr12:6937342-6937462;chr12:6937461-6937581;chr12:6937581-6937701;chr12:6937700-6937820;chr12:6937819-6937939;chr12:6937939-6938059;chr12:6938058-6938178;chr12:6938178-6938298;chr12:6938297-6938417;chr12:6938416-6938536;chr12:6938536-6938656;chr12:6938655-6938775;chr12:50505053-50505173;chr12:50505169-50505289;chr12:50505286-50505406;chr12:50505402-50505522;chr12:50505523-50505643;chr12:50505639-50505759;chr12:50505756-50505876;chr12:50505872-50505992;chr12:50505993-50506113;chr12:50506109-; 50506229;chr12:50506226-50506346;chr12:50506342-50506462;chr12:50506463-50506583;chr12:50506579-50506699;chr12:50506696-50506816;chr12:50506812-50506932;chr12:50506933-50507053;chr12:111596949-111597069;chr12:111597065-111597185;chr12:111597182-111597302;chr12:111597298-111597418;chr12:111597419-111597539;chr12:111597535-111597655;chr12:111597652-111597772;chr12:111597768-111597888;chr12:111597889-111598009;chr12:111598005-111598125;chr12:111598122-111598242;chr12:111598238-111598358;chr12:111598359-111598479;chr12:111598475-111598595;chr12:111598592-111598712;chr12:111598708-111598828;chr12:111598829-111598949;chr12:111599019-111599139;chr12:111599135-111599255;chr12:111599252-111599372;chr12:111599368-111599488;chr12:111599489-111599609;chr12:111599605-111599725;chr12:111599722-111599842;chr12:111599838-111599958;chr12:111599959-111600079;chr12:111600075-111600195;chr12:111600192-111600312;chr12:111600308-111600428;chr12:111600429-111600549;chr12:111600545-111600665;<h2 style=";text-align:left;direction:ltr">chr12:111600662-111600782;chr13:70137383-70137503;chr13:70137501-70137621;chr13:70137620-70137740;chr13:70138214-70138334;chr13:70138453-70138573;chr13:70138572-70138692;chr13:70138690-70138810;chr13:70138811-70138931;chr13:70138929-7 0139049;chr13:70139048-70139168;chr13:70139166-70139286;chr13:70139287-70139407;chr13:70139523-70139643;chr13:7013964 1-70139761;chr13:70139762-70139882;chr13:70139880-70140000;chr13:70139999-70140119;chr13:70140117-70140237;chr13:70140 238-70140358;chr13:70140356-70140476;chr13:70140475-70140595;chr13:70140593-70140713;chr13:70140714-70140834;chr13:70140832-70140952;chr13:70140951-70141071;chr13:70141069-70141189;chr13:70141190-70141310;chr13:70141309-70141429;chr13 :99983448-99983568;chr13:99983566-99983686;chr13:99983685- 99983805;chr13:99983803-99983923;chr13:99983924-99984044;ch r13:99984042-99984162;chr13:99984161-99984281;chr13:99984279-99984399;chr13:99984400-99984520;chr13:99984518-99984638; chr13:99984637-99984757;chr13:99984755-99984875;chr13:99984876-99984996;chr13:99984994-99985114;chr13:99985113-99985233;chr13:99985231-99985351;chr13:99985352-99985472;chr13:99985470-99985590;chr13:99985588-99985708;chr13:99985706-99985826;chr13:99985945-99986065;chr13:99986064-99986184;chr13:99986182-99986302;chr13:99986303-99986423;chr13:99986421-99986541;chr13:99986540-99986660;chr13:99986658-99986778;chr13:99986779-99986899;chr13:99986897-99987017;chr13:99987016-99987136;chr13:99987134-99987254;chr13:99987255-99987375;chr13:99987374-99987494;chr14:23319463-23319583;chr14:23319579-23319699;chr14:23319696-23319816;chr14:23319812-23319932;chr14:23319933-23320053;chr14:23320049-23320169;chr14:23320166-23320286;chr14:23320282-23320402;chr14:23320403-23320523;chr14:23320519-23320639;chr14:23320636-23320756;chr14:23320752-23320872;chr14:23320873-23320993;chr14:23320989-23321109;chr14:23321106-23321226;chr14:23321222-23321342;chr14:23321343-23321463;chr14:23321543-23321663;chr14:23321659-23321779;chr14:23321776-23321896;chr14:23321892-23322012;chr14:23322013-23322133;chr14:23322129-23322 249. 322836;chr14:23322832-23322952;chr14:23322953-23323073;chr14:23323069-23323189;chr14:23323186-23323306;chr14:2332330 2-23323422;chr14:23323423-23323543;chr14:92071052-92071172;chr14:92071168-92071288;chr14:92071638-92071758;chr14:9207 1755-92071875;chr14:92071871-92071991;chr14:92071992-92072112;chr14:92072108-92072228;chr14:92072225-92072345;chr14: 92072341-92072461;chr14:92072462-92072582;chr14:92072811-92072931;chr14:92072932-92073052;chr15:22786703-22786823;chr 15:22786819-22786939;chr15:22786936-22787056;chr15:22787052-22787172;chr15:22787173-22787293;chr15:22787289-22787409 ;chr15:22787406-22787526;chr15:22787522-22787642;chr15:22787643-22787763;chr15:22787759-22787879;chr15:22787876-2278; 7996;chr15:22787992-22788112;chr15:22788113-22788233;chr15:22788583-22788703;chr16:17468869-17468989;chr16:17468985-1746910 5;chr16:17469102-17469222;chr16:17469218-17469338;chr16:174693 39-17469459;chr16:17469455-17469575;chr16:17469572-17469692;ch r16:17469688-17469808;chr16:17469809-17469929;chr16:17469925-17470045;chr16:17470042-17470162;chr16:17470158-17470278;chr16 :17470279-17470399;chr16:17470395-17470515;chr16:17470512-17470632;chr16:17470628-17470748;chr16:17470749-17470869;chr16:174 71083-17471203;chr16:17471200-17471320;chr16:17471316-17471436;chr16:17471437-17471557;chr16:17471553-17471673;chr16:174716 70-17471790;chr16:17471786-17471906;chr16:17471907-17472027;chr16:17472023-17472143;chr16:17472140-17472260;chr16:17472256-1 7472376;chr16:17472847-17472967;chr16:24611438-24611558;chr16:24611554-24611674;chr16:24611671-24611791;chr16:24612141-24612 261;chr16:24612257-24612377;chr16:24612378-24612498;chr16:24612727-24612847;chr16:24612848-24612968;chr16:24613197-24613317;chr16:24613318-24613438;chr16:24613881-24614001;chr16:24614002-24614122;chr16:24614118-24614238;chr16:24614235-24614355;chr16:24614351-24614471;chr16:24614472-24614592;chr16:24614588-24614708;chr16:24614705-24614825;chr16:24615058- 24615178;chr16:24615175-24615295;chr16:24615291-24615411;chr16:24615412-24615532;chr16:66487396-66487516;chr16:66487514-66487634;chr16:66487633-66487753;chr16:66487751-66487871;chr16:66487872-66487992;chr16:66487990-66488110;chr16:6 66488109-66488229;chr16:66488227-66488347;chr16:66489181-66489301;chr16:66489299-66489419;chr16:66489420-66489540;chr16:66489538-66489658;chr16:66489657-66489777;chr16:66489775-66489895;chr16:66489896-66490016;chr16:66490014-66490134 ;chr16:66490490-66490610;chr16:66490609-66490729;chr16:66490727-66490847;chr16:66490848-66490968;chr16:66490966-6649 1086;chr16:66491085-66491205;chr16:66491203-66491323;chr16:66491324-66491444;chr16:66491442-66491562;chr16:66491920; -66492040;chr16:66492275-66492395;chr16:66492751-66492871;chr16:66492872-66492992;chr16:66492990-66493110;chr16:66493109-664 93229;chr16:66493227-66493347;chr16:66493348-66493468;chr16:87602282-87602402;chr16:87602637-87602757;chr16:87602758-8760287 8;chr16:87602876-87602996;chr16:87602995-87603115;chr16:876031 13-87603233;chr16:87603234-87603354;chr16:87603352-87603472;ch r16:87603471-87603591;chr16:87603589-87603709;chr16:87603710-87603830;chr16:87603828-87603948;chr16:87603947-87604067;chr16:8 7604065-87604185;chr16:87604186-87604306;chr16:87604304-87604424;chr16:87604423-87604543;chr16:87604541-87604661;chr16:87604 662-87604782;chr16:87604780-87604900;chr16:87604899-87605019;chr16:87605017-87605137;chr16:87605138-87605258;chr16:87605256- 87605376;chr16:87605375-87605495;chr16:87605493-87605613;chr16:87605614-87605734;chr16:87605732-87605852;chr16:87605851-8760 5971;chr16:87605969-87606089;chr16:87606090-87606210;chr16:876 06209-87606329;chr17:80145001-80145121;chr17:80145117-80145237;chr17:80145587-80145707;chr17:80145704-80145824;chr17:80145820-80145940;chr17:80145941-80146061;chr17:80146057-80146177;chr17:80146174-80146294;chr17:80146290-80146410;chr17:80146411-80146531;chr17:80146527-80146647;chr17:80146644-80146764;chr17:80146760-80146880;chr17:80146881-80147001;chr18:55584113-55584233;chr18:55584229-55584349;chr18:55584346-55584466;chr18:55584462-55584582;chr18:55584583-55584703;chr18:55584699-55584819;chr18:55584816-55584936;chr18:55584932-55585052;chr18:55585053-55585173;chr18:55585169-55585289;chr18:55585286-55585406;chr18:55585402-55585522;chr18:55585523-55585643;chr18:55585639-55585759;chr18:55585756-55585876;chr18:55585872-55585992;chr18:55585993-55586113;chr18:55586229-55586349;chr18:55586345-55586465;chr18:55586462-55586582;chr18:55586578-55586698;chr18:55586699-55586819;chr18:55586815-55586935;chr18:55586932-55587052;chr18:55587048-55587168;chr18:55587402-55587522;chr18:5558; 7518-55587638;chr18:55587639-55587759;chr18:55587755-55587875;chr18:55587872-55587992;chr18:55587988-55588108;chr18:55588109-55588229;chr18:57022495-57022615;chr18:57022611-57022731;chr18:57022728-57022848;chr18:57022844-57022964;chr18:57022965-57023085;chr18:57023081-57023201;chr18:57023198-57023318;chr18:57023314-57023434;chr18:57023435-57023555;chr18:57023551-57023671;chr18:57023668-57023788;chr18:57023784-57023904;chr18:57023905-57024025;chr18:57024021-57024141;chr18:57024138-57024258;chr18:57024254-57024374;chr18:57024375-57024495;chr18:57024955-57025075;chr18:57025071-57025191;chr18:57025188-57025308;chr18:57025304-57025424;chr18:57025425-57025545;chr18:57025541-57025661;chr18:57025658-57025778;chr18:57025774-57025894;chr18:57025895-57026015;chr18:57026011-57026131;chr18:57026128-57026248;chr18:57026244-57026364;chr18:57026365-57026485;chr18:57026481-57026601;chr18:57026598-57026718;chr18:57026714-57026834;chr18:57026835-57026955;chr18:57024494-57024614;chr18:57024572-57024692;<h2 style=";text-align:left;direction:ltr">chr18:57024650-57024770;chr19:1047438-1047558;chr19:1047554-1047674;chr19:1047671-1047791;chr19:1047787-1047907;chr19:1048848-1048968;chr19:1048964-1049084;chr19:1049081-1049201;chr19:1049197-1049317;chr19:1049318-1049438;chr19:1 050841-1050961;chr19:1050962-1051082;chr19:1051078-1051198;chr19:1051195-1051315;chr19:1051311-1051431;chr19:1051432-1051552;chr19:1051548-1051668;chr19:1051665-1051785;chr19:1051781-1051901;chr19:1051902-1052022;chr19:1046437-104 6557;chr19:1046543-1046663;chr19:1046650-1046770;chr19:1046756-1046876;chr19:1046877-1046997;chr19:1046983-1047103;chr19:1047090-1047210;chr19:1047196-1047316;chr19:1047317-1047437;chr19:1052023-1052143;chr19:1052129-1052249;chr19 :1052236-1052356;chr19:1052782-1052902;chr19:13206333-13206453;chr19:13206451-13206571;chr19:13206569-13206689;chr19:13206687-13206807;chr19:13206808-13206928;chr19:13206926-13207046;chr19:13207044-13207164;chr19:13207162-1320728; <h2 style=";text-align:left;direction:ltr">2;chr19:13207283-13207403;chr19:13207401-13207521;chr19:13207520-13207640;chr19:13207638-13207758;chr19:13207759-13207879;c hr19:13207877-13207997;chr19:13207995-13208115;chr19:13208234-13208354;chr19:13208352-13208472;chr19:13208470-13208590;chr1 9:13208588-13208708;chr19:13208709-13208829;chr19:13208827-13208947;chr19:13208945-13209065;chr19:13209063-13209183;chr19:1 3209184-13209304;chr19:13209302-13209422;chr19:13209420-13209540;chr19:13209538-13209658;chr19:13209659-13209779;chr19:13209 778-13209898;chr19:14494396-14494516;chr19:14494635-14494755;chr19:14494753-14494873;chr19:14494871-14494991;chr19:14494992 -14495112;chr19:14495110-14495230;chr19:14495229-14495349;chr19:14495347-14495467;chr19:14495468-14495588;chr19:14495586-14 495706;chr19:14495705-14495825;chr19:14495823-14495943;chr19:14495944-14496064;chr19:14496062-14496182;chr19:14496180-14496 300;chr19:14496298-14496418;chr19:14496419-14496539;chr19:14496537-14496657;chr19:14496656-14496776;chr19:14496774-14496894;<h2 style=";text-align:left;direction:ltr">chr19:14496895-14497015;chr19:14497013-14497133;chr19:14497131-14497251;chr19:14497249-14497369;chr19:14497370-14497490;chr19:14497488-14497608;chr19:14497607-14497727;chr19:14497725-14497845;chr19:18784027-18784147;chr19:18784144-1 8784264;chr19:18784262-18784382;chr19:18784379-18784499;chr19:18784500-18784620;chr19:18784617-18784737;chr19:18784735-18784855;chr19:18784852-18784972;chr19:18784973-18785093;chr19:18785090-18785210;chr19:18785208-18785328;chr19:187 85325-18785445;chr19:18785446-18785566;chr19:18785563-18785683;chr19:18785681-18785801;chr19:18785798-18785918;chr19:18785919-18786039;chr19:18786036-18786156;chr19:18786154-18786274;chr19:18786271-18786391;chr19:18786392-18786512;c hr19:18786510-18786630;chr19:18786627-18786747;chr19:18787101-18787221;chr19:18787218-18787338;chr19:18787339-18787459;chr19:18787456-18787576;chr19:18787574-18787694;chr19:18787691-18787811;chr19:45768204-45768324;chr19:45768320-45; <h2 style=";text-align:left;direction:ltr">768440;chr19:45768437-45768557;chr19:45768553-45768673;chr19:45768674-45768794;chr19:45768790-45768910;chr19:45768907-4576902 7;chr19:45769023-45769143;chr19:45769144-45769264;chr19:45769260-45769380;chr19:45769377-45769497;chr19:45769493-45769613;chr 19:45769614-45769734;chr19:45769730-45769850;chr19:45769847-45769967;chr19:45769963-45770083;chr19:45770084-45770204;chr19:45 770266-45770386;chr19:45770382-45770502;chr19:45770499-45770619;chr19:45770615-45770735;chr19:45770736-45770856;chr19:4577085 2-45770972;chr19:45770969-45771089;chr19:45771085-45771205;chr19:45771206-45771326;chr19:45771322-45771442;chr19:45771439-457 71559;chr19:45771555-45771675;chr19:45771676-45771796;chr19:45771792-45771912;chr19:45771909-45772029;chr19:45772025-45772145 ;chr19:45772146-45772266;chr2:100102799-100102919;chr2:100102916-100103036;chr2:100103034-100103154;chr2:100103151-100103271; chr2:100103272-100103392;chr2:100103390-100103510;chr2:1001035 07-100103627;chr2:100103625-100103745;chr2:100103746-100103866;chr2:100103863-100103983;chr2:100103981-100104101;chr2:100104098-100104218;chr2:100104219-100104339;chr2:100104336-100104456;chr2:100104454-100104574;chr2:100104571-100104691;chr2:100104810-100104930;chr2:100104927-100105047;chr2:100105045-100105165;chr2:100105166-100105286;chr2:100105283-100105403;chr2:100105401-100105521;chr2:100105518-100105638;chr2:100105639-100105759;chr2:100105756-100105876;chr2:100105874-100105994;chr2:100105991-100106111;chr2:100106112-100106232;chr2:100106230-100106350;chr2:100106347-100106467;chr2:100106465-100106585;chr2:100106586-100106706;chr2:100106704-100106824;chr2:176091057-176091177;chr2:176091175-176091295;chr2:176091293-176091413;chr2:176091411-176091531;chr2:176091532-176091652;chr2:176091650-176091770;chr2:176091769-176091889;chr2:176091887-176092007;chr2:176092008-176092128;chr2:176092126-176092246;chr2:176092244-176092364;chr2:176092362-176092482;chr2:17609; 2483-176092603;chr2:176092601-176092721;chr2:176092720-176092840;chr2:176092838-176092958;chr2:176092959-176093079;chr2:176093077-176093197;chr2:176093195-176093315;chr2:176093313-176093433;chr2:176093434-176093554;chr2:176093552-176093672;chr2:176093670-176093790;chr2:176093788-176093908;chr2:176093909-176094029;chr2:176094027-176094147;chr2:176094146-176094266;chr2:176094264-176094384;chr2:176094385-176094505;chr2:176094503-176094623;chr2:176094621-176094741;chr2:176094739-176094859;chr2:176094860-176094980;chr2:176094979-176095099;chr2:190878872-190878992;chr2:190878990-190879110;chr2:190879109-190879229;chr2:190879227-190879347;chr2:190879348-190879468;chr2:190879703-190879823;chr2:190879824-190879944;chr2:190879942-190880062;chr2:190880061-190880181;chr2:190880179-190880299;chr2:190880300-190880420;chr2:190880418-190880538;chr2:190880537-190880657;chr2:190880655-190880775;chr2:190880776-190880896;chr2:190880896-190881016;chr2:190881014-190881134;chr2:190881133-190881253;<h2 style=";text-align:left;direction:ltr">chr2:190881251-190881371;chr2:190881372-190881492;chr2:19 0881490-190881610;chr2:190881609-190881729;chr2:190881727- 190881847;chr2:190881848-190881968;chr2:190881966-1908820 86;chr2:190882085-190882205;chr2:190882203-190882323;chr2: 190882324-190882444;chr2:190882442-190882562;chr2:190882561-190882681;chr2:190882679-190882799;chr2:190882800-190882920;chr20:2650732-2650852;chr20:2651208-2651328;chr20:2651326-2651446;chr20:2651444-2651564;chr20:2651562-2651682; chr20:2651683-2651803;chr20:2651801-2651921;chr20:2651920-2652040;chr20:2652038-2652158;chr20:2652159-2652279;chr20:2652277-2652397;chr20:2652395-2652515;chr20:2652513-2652633;chr20:2652634-2652754;chr20:2652752-2652872;chr20:2652 871-2652991;chr20:2652989-2653109;chr20:2653110-2653230;chr20:2653228-2653348;chr20:2653346-2653466;chr20:2653464-2653584;chr20:2653585-2653705;chr20:2653940-2654060;chr20:2654061-2654181;chr20:2654179-2654299;chr20:2654297-265441; <h2 style=";text-align:left;direction:ltr">7;chr20:2654415-2654535;chr20:2654536-2654656;chr20:2654655-2654775;chr20:4697398-4697518;chr20:4697516-4697636;chr20:46976 35-4697755;chr20:4697753-4697873;chr20:4697874-4697994;chr20: 4697992-4698112;chr20:4698111-4698231;chr20:4698229-4698349;ch r20:4698350-4698470;chr20:4698587-4698707;chr20:4698705-4698825;chr20:4698826-4698946;chr20:4698944-4699064;chr20:4699063-4 699183;chr20:4699181-4699301;chr20:4699302-4699422;chr20:4699422-4699542;chr20:4699540-4699660;chr20:4699659-4699779;chr20:4 699777-4699897;chr20:4699898-4700018;chr20:4700016-4700136;chr20:4700135-4700255;chr20:4700253-4700373;chr20:4700374-470049 4;chr20:4700492-4700612;chr20:4700611-4700731;chr20:4700729-4700849;chr20:4700850-4700970;chr20:4700968-4701088;chr20:47010 87-4701207;chr20:4701205-4701325;chr20:4701326-4701446;chr21: 43774444-43774564;chr21:43774560-43774680;chr21:43774677-43774 797;chr21:43774793-43774913;chr21:43775263-43775383;chr21:4377 5384-43775504;chr21:43775500-43775620;chr21:43775617-43775737;<h2 style=";text-align:left;direction:ltr">chr21:43775733-43775853;chr21:43775854-43775974;chr21:43775970-43776090;chr21:43776087-43776207;chr21:43776203-43776323;chr21:43776324-43776444;chr22:19764762-19764882;chr22:19764880-19765000;chr22:19764999-19765119;chr22:19765117-1 9765237;chr22:19765238-19765358;chr22:19765356-19765476;chr22:19765475-19765595;chr22:19765593-19765713;chr22:19765714-19765834;chr22:19765832-19765952;chr22:19765950-19766070;chr22:19766068-19766188;chr22:19766189-19766309;chr22:19 766307-19766427;chr22:19766426-19766546;chr22:19766544-19766664;chr22:19766665-19766785;chr22:19766783-19766903;chr22:19766902-19767022;chr22:19767020-19767140;chr22:19767141-19767261;chr22:19767259-19767379;chr22:19767378-19767498; chr22:19767496-19767616;chr22:19767617-19767737;chr22:19767735-19767855;chr22:19767853-19767973;chr22:19767971-19768091;chr22:19768092-19768212;chr22:19768210-19768330;chr22:19768329-19768449;chr22:19768447-19768567;chr22:19768568-1; <h2 style=";text-align:left;direction:ltr">9768688;chr22:19768687-19768807;chr22:45793354-45793474;chr22:45793470-45793590;chr22:45793587-45793707;chr22:45793703-4579 3823;chr22:45793824-45793944;chr22:45794173-45794293;chr22:45794410-45794530;chr22:45794527-45794647;chr22:45794643-45794763 ;chr22:45794764-45794884;chr22:45794880-45795000;chr22:45794997-45795117;chr22:45795113-45795233;chr22:45795234-45795354;chr 22:45795330-45795450;chr3:63910684-63910804;chr3:63910802-6391 0922;chr3:63910919-63911039;chr3:63911037-63911157;chr3:639111 58-63911278;chr3:63911276-63911396;chr3:63911393-63911513;chr 3:63911511-63911631;chr3:63911632-63911752;chr3:63911750-63911 870;chr3:63911868-63911988;chr3:63911986-63912106;chr3:6391210 7-63912227;chr3:63912225-63912345;chr3:63912342-63912462;chr3: 63912460-63912580;chr3:63912581-63912701;chr3:63912699-6391281 9;chr3:63912816-63912936;chr3:63912934-63913054;chr3:63913055- 63913175;chr3:63913173-63913293;chr3:63913290-63913410;chr3:63 913408-63913528;chr3:63913529-63913649;chr3:63913647-63913767;chr3:63913764-63913884;chr3:63913882-63914002;chr3:63914003-63914123;chr3:63914121-63914241;chr3:63914238-63914358;chr3:63914356-63914476;chr3:63914477-63914597;chr3:63914596-63914716;chr3:129170576-129170696;chr3:129170692-129170812;chr3:129170809-129170929;chr3:129170925-129171045;chr3:129171046-129171166;chr3:129171162-129171282;chr3:129171279-129171399;chr3:129171395-129171515;chr3:129171516-129171636;chr3:129171632-129171752;chr3:129171749-129171869;chr3:129172219-129172339;chr3:129172775-129172895;chr3:129172892-129173012;chr3:129173008-129173128;chr3:129173245-129173365;chr3:129173362-129173482;chr3:129173478-129173598;chr3:129173599-129173719;chr3:129173715-129173835;chr3:129173832-129173952;chr3:129173948-129174068;chr3:129174069-129174189;chr3:129174185-129174305;chr3:138944021-138944141;chr3:138944139-138944259;chr3:138944257-138944377;chr3:138944375-138944495;chr3:138944496-138944616;chr3:138944614-1; 38944734;chr3:138944733-138944853;chr3:138944851-138944971;chr3:138944972-138945092;chr3:138945090-138945210;chr3:138945208-138945328;chr3:138945326-138945446;chr3:138945447-138945567;chr3:138945565-138945685;chr3:138945683-138945803;chr3:138945801-138945921;chr3:138945922-138946042;chr3:138946040-138946160;chr3:138946158-138946278;chr3:138946276-138946396;chr3:138946397-138946517;chr3:138946515-138946635;chr3:138946634-138946754;chr3:138946752-138946872;chr3:138946873-138946993;chr3:138946991-138947111;chr3:138947109-138947229;chr3:138947227-138947347;chr3:138947348-138947468;chr3:138947466-138947586;chr3:138947584-138947704;chr3:138947702-138947822;chr3:138947823-138947943;chr3:138947942-138948062;chr3:183710176-183710296;chr3:183710292-183710412;chr3:183710409-183710529;chr3:183710525-183710645;chr3:183710646-183710766;chr3:183710762-183710882;chr3:183710879-183710999;chr3:183710995-183711115;chr3:183711116-183711236;chr3:183711465-183711585;chr3:183711586-183711706;chr3:183711702-183711822;chr4:3073109-3073229;chr4:3073225-3073345;chr4:3073346-3073466;chr4:3073462-3073582;chr4:3073579-3073699;chr4:3073695-3073815;chr4:3073816-3073936;chr4:3073932-3074052;chr4:3074049-3074169;chr4:3074165-3074285;chr4:3074286-3074406;chr4:3074402-3074522;chr4:3074519-3074639;chr4:3074635-3074755;chr4:3074756-3074876;chr4:3074940-3075060;chr4:3075056-3075176;chr4:3075173-3075293;chr4:3075289-3075409;chr4:3075410-3075530;chr4:3075526-3075646;chr4:3075643-3075763;chr4:3075759-3075879;chr4:3075880-3076000;chr4:3075996-3076116;chr4:3076113-3076233;chr4:3076229-3076349;chr4:3076350-3076470;chr4:3076466-3076586;chr4:3076583-3076703;chr4:3076699-3076819;chr4:39346543-39346663;chr4:39346663-39346783;chr4:39346782-39346902;chr4:39346901-39347021;chr4:39347021-39347141;chr4:39347140-39347260;chr4:39347260-39347380;chr4:39347379-39347499;chr4:39347618-39347738;chr4:39347737-39347857;chr4:39347856-39347976;chr4:; 39347976-39348096;chr4:39348453-39348573;chr4:39348573-39348693;chr4:39348692-39348812;chr4:39348811-39348931;chr4:39348931-39349051;chr4:39349050-39349170;chr4:39349289-39349409;chr4:39349408-39349528;chr4:39349527-39349647;chr4:39349647-39349767;chr4:39350124-39350244;chr4:39350244-39350364;chr4:39350363-39350483;chr4:41744093-41744213;chr4:41744212-41744332;chr4:41744330-41744450;chr4:41744451-41744571;chr4:41744569-41744689;chr4:41744688-41744808;chr4:41744806-41744926;chr4:41744927-41745047;chr4:41745045-41745165;chr4:41745164-41745284;chr4:41745403-41745523;chr4:41745521-41745641;chr4:41745640-41745760;chr4:41745758-41745878;chr4:41745879-41745999;chr4:41745997-41746117;chr4:41746116-41746236;chr4:41746234-41746354;chr4:41746355-41746475;chr4:41746473-41746593;chr4:41746592-41746712;chr4:41746710-41746830;chr4:41746831-41746951;chr4:41746949-41747069;chr4:41747068-41747188;chr4:41747186-41747306;chr4:41747307-41747427;chr4:41747425-41747545;chr4:41747544-41747664;chr4:41747662-41747782;chr4:41747783-41747903;chr4:41747902-41748022;chr4:159340526-159340646;chr4:159340875-159340995;chr4:159340996-159341116;chr4:159341112-159341232;chr4:159341229-159341349;chr4:159341345-159341465;chr4:159341466-159341586;chr4:159341582-159341702;chr4:159341699-159341819;chr4:159341815-159341935;chr4:159341936-159342056;chr4:159342052-159342172;chr4:159342169-159342289;chr4:159342285-159342405;chr4:159342406-159342526;chr4:159342618-159342738;chr4:159342734-159342854;chr4:159342851-159342971;chr4:159342967-159343087;chr4:159343088-159343208;chr4:159343204-159343324;chr4:159343321-159343441;chr4:159343437-159343557;chr4:159343558-159343678;chr4:159343674-159343794;chr4:159343791-159343911;chr4:159343907-159344027;chr4:159344028-159344148;chr4:159344144-159344264;chr4:159344261-159344381;chr4:159344377-159344497;chr4:159344498-159344618;chr5:10354338-10354458;chr5:10354454-10354574;chr5:10354571; <h2 style=";text-align:left;direction:ltr">-10354691;chr5:10354687-10354807;chr5:10354808-10354928;chr5:10354924-10355044;chr5:10355041-10355161;chr5:10355157-103552 77;chr5:10355278-10355398;chr5:10355394-10355514;chr5:10355511-10355631;chr5:10355627-10355747;chr5:10355748-10355868;chr5: 10355864-10355984;chr5:10355981-10356101;chr5:10356097-10356217;chr5:10356218-10356338;chr5:10356760-10356880;chr5:1035688 1-10357001;chr5:10356997-10357117;chr5:10357114-10357234;chr5:10357230-10357350;chr5:10357351-10357471;chr5:10357467-103575 87;chr5:10357584-10357704;chr5:10357700-10357820;chr5:10357821-10357941;chr5:10358054-10358174;chr5:10358170-10358290;chr5 :10358291-10358411;chr5:146876727-146876847;chr5:146876845-146876965;chr5:146876962-146877082;chr5:146877080-146877200;chr5 :146877201-146877321;chr5:146877319-146877439;chr5:146877436- 146877556;chr5:146877554-146877674;chr5:146877675-146877795;ch r5:146877793-146877913;chr5:146877911-146878031;chr5:146878029-146878149;chr5:146878150-146878270;chr5:146878268-146878388;chr5:146878385-146878505;chr5:146878503-146878623;chr5:146878624-146878744;chr5:146878742-146878862;chr5:146878859-1 46878979;chr5:146878977-146879097;chr5:146879098-146879218;chr5:146879216-146879336;chr5:146879333-146879453;chr5:146879336 879451-146879571;chr5:146879572-146879692;chr5:146879690-146879810;chr5:146879807-146879927;chr5:146879925-146880045 ;chr5:146880046-146880166;chr5:146880281-146880401;chr5:146880399-146880519;chr5:146880520-146880640;chr5:146880639-1 46880759;chr6:16325633-16325753;chr6:16325982-16326102;chr6:16326103-16326223;chr6:16326219-16326339;chr6:16326336-16326456;chr6:16326452-16326572;chr6:16326573-16326693;chr6:16326689-16326809;chr6:16326806-16326926;chr6:16326922-163 27042;chr6:16327043-16327163;chr6:16327159-16327279;chr6:16327276-16327396;chr6:16327392-16327512;chr6:16327513-16327633;chr6:16327724-16327844;chr6:16327840-16327960;chr6:16327957-16328077;chr6:16328073-16328193;chr6:16328194-163283; 14;chr6:16328310-16328430;chr6:16328427-16328547;chr6:16328543-16328663;chr6:16329013-16329133;chr6:16329134-16329254;chr6:16329250-16329370;chr6:16329367-16329487;chr6:16329483-16329603;chr6:16329604-16329724;chr6:45420678-45420798;chr6:45420794-4 5420914;chr6:45420911-45421031;chr6:45421027-45421147;chr6:45421148-45421268;chr6:45421264-45421384;chr6:45421381-45421501;chr6:45421497-45421617;chr6:45421618-45421738;chr6:45421734-45421854;chr6:45421851-45421971;chr6:45421967-45422087;chr6:45422 088-45422208;chr6:45422204-45422324;chr6:45422321-45422441;chr6:45422437-45422557;chr6:45422558-45422678;chr6:45422801-45422921;chr6:45422917-45423037;chr6:45423034-45423154;chr6:45423150-45423270;chr6:45423271-45423391;chr6:45423387-45423507;chr6: 45423504-45423624;chr6:45423620-45423740;chr6:45423741-45423861;chr6:45423857-45423977;chr6:45423974-45424094;chr6:45424090-45424210;chr6:45424211-45424331;chr6:45424327-45424447;chr6:4542444-45424564;chr6:45424560-45424680;chr6:45424681-45424801;<h2 style=";text-align:left;direction:ltr">chr6:170562017-170562137;chr6:170562133-170562253;chr6:17 0562250-170562370;chr6:170562366-170562486;chr6:170562487- 170562607;chr6:170562603-170562723;chr6:170562720-1705628 40;chr6:170562836-170562956;chr6:170562957-170563077;chr6: 170563073-170563193;chr6:170563190-170563310;chr6:170563306-170563426;chr6:170563427-170563547;chr6:170563543-170563663;chr6:170563660-170563780;chr6:170563776-170563896;chr6:170563897-170564017;chr7:27197678-27197798;chr7:27197794 -27197914;chr7:27197911-27198031;chr7:27198027-27198147;chr7:27198148-27198268;chr7:27198264-27198384;chr7:27198381-27198501;chr7:27198497-27198617;chr7:27198618-27198738;chr7:27198734-27198854;chr7:27198851-27198971;chr7:27198967- 27199087;chr7:27199088-27199208;chr7:27199204-27199324;chr7:27199321-27199441;chr7:27199437-27199557;chr7:27199558-27199678;chr7:27199967-27200087;chr7:27200083-27200203;chr7:27200200-27200320;chr7:27200316-27200436;chr7:27200437-; <h2 style=";text-align:left;direction:ltr">27200557;chr7:27200553-27200673;chr7:27200670-27200790;chr7:27200786-27200906;chr7:27200907-27201027;chr7:27201023-27201143 ;chr7:27201140-27201260;chr7:27201256-27201376;chr7:27201377-27201497;chr7:27201493-27201613;chr7:27201610-27201730;chr7:27 201726-27201846;chr7:27201847-27201967;chr8:104586964-104587084;chr8:104587082-104587202;chr8:104587200-104587320;chr8:1045 87318-104587438;chr8:104587439-104587559;chr8:104587557-10458 7677;chr8:104587674-104587794;chr8:104587792-104587912;chr8:10 4587913-104588033;chr8:104588031-104588151;chr8:104588149-104 588269;chr8:104588267-104588387;chr8:104588388-104588508;chr8: 104588506-104588626;chr8:104588623-104588743;chr8:104588741-1 04588861;chr8:104589098-104589218;chr8:104589216-104589336;chr 8:104589337-104589457;chr8:104589455-104589575;chr8:104589572 -104589692;chr8:104589690-104589810;chr8:104589811-104589931;c hr8:104589929-104590049;chr8:104590047-104590167;chr8:104590165-104590285;chr8:104590286-104590406;chr8:104590760-104590880;<h2 style=";text-align:left;direction:ltr">chr8:104590879-104590999;chr8:118364812-118364932;chr8:118 364928-118365048;chr8:118365045-118365165;chr8:118365161-1 18365281;chr8:118365398-118365518;chr8:118365515-118365635 ;chr8:118365752-118365872;chr8:118365868-118365988;chr8:118 365985-118366105;chr8:118366101-118366221;chr8:118366222-1 18366342;chr8:118366338-118366458;chr8:118366918-118367038 ;chr8:118367034-118367154;chr8:118367151-118367271;chr8:11 8367267-118367387;chr8:118367388-118367508;chr8:118367504-1 18367624;chr8:118367621-118367741;chr8:118367737-118367857 ;chr8:118367858-118367978;chr8:118367974-118368094;chr8:11 8368091-118368211;chr8:118368207-118368327;chr8:118368328- 118368448;chr8:118368444-118368564;chr8:118368561-118368681 ;chr8:118368677-118368797;chr8:118368798-118368918;chr9:27571717-27571837;chr9:27571833-27571953;chr9:27571954-275720 74;chr9:27572070-27572190;chr9:27572187-27572307;chr9:27572303-27572423;chr9:27572424-27572544;chr9:27572540-2757266; <h2 style=";text-align:left;direction:ltr">0;chr9:27572657-27572777;chr9:27572773-27572893;chr9:27572894-27573014;chr9:27573010-27573130;chr9:27573127-27573247;chr9:275 73243-27573363;chr9:27573364-27573484;chr9:27573546-27573666;chr9:27573662-27573782;chr9:27573779-27573899;chr9:27573895-2757 4015;chr9:27574016-27574136;chr9:27574132-27574252;chr9:27574249-27574369;chr9:27574365-27574485;chr9:27574486-27574606;chr9: 27574602-27574722;chr9:27574719-27574839;chr9:27574835-27574955;chr9:27574956-27575076;chr9:27575072-27575192;chr9:27575189-2 7575309;chr9:27575426-27575546;chr9:69035275-69035395;chr9:69035391-69035511;chr9:69035508-69035628;chr9:69035624-69035744;ch r9:69035745-69035865;chr9:69035861-69035981;chr9:69035978-69036098;chr9:69036094-69036214;chr9:69036215-69036335;chr9:6903644 8-69036568;chr9:69036564-69036684;chr9:69036685-69036805;chr9:69036801-69036921;chr9:69036918-69037038;chr9:69037034-69037154 ;chr9:130681641-130681761;chr9:130681757-130681877;chr9:130681874-130681994;chr9:130681990-130682110;chr9:130682111-130682231;chr9:130682227-130682347;chr9:130682344-130682464;chr9:130682460-130682580;chr9:130682581-130682701;chr9:130682697-130682817;chr9:130682814-130682934;chr9:130682930-130683050;chr9:130683051-130683171;chr9:130683167-130683287;chr9:130683284-130683404;chr9:130683400-130683520;chr9:130683521-130683641;chrX:25011529-25011649;chrX:25011645-25011765;chrX:25011762-25011882;chrX:25011878-25011998;chrX:25011999-25012119;chrX:25012115-25012235;chrX:25012232-25012352;chrX:25012348-25012468;chrX:25012469-25012589;chrX:25012585-25012705;chrX:25012702-25012822;chrX:25012818-25012938;chrX:25012939-25013059;chrX:25013055-25013175;chrX:25013172-25013292;chrX:25013288-25013408;chrX:25013409-25013529;chrX:25013697-25013817;chrX:25013813-25013933;chrX:25014283-25014403;chrX:25014400-25014520;chrX:25014516-25014636;chrX:25014637-25014757;chrX:25014753-25014873;chrX:25014870-25014990;chrX:25014986-25015106;chrX:; 25015107-25015227;chrX:25015223-25015343;chrX:25015340-25015460;chrX:25015456-25015576;chrX:25015577-25015697;chrX:31282557-31282677;chrX:31282675-31282795;chrX:31282794-31282914;chrX:31282912-31283032;chrX:31283033-31283153;chrX:31283152-31283272;chrX:31283272-31283392;chrX:31283390-31283510;chrX:31283509-31283629;chrX:31283627-31283747;chrX:31283748-31283868;chrX:31283866-31283986;chrX:31284343-31284463;chrX:31284462-31284582;chrX:31284700-31284820;chrX:31284819-31284939;chrX:31284937-31285057;chrX:31285058-31285178;chrX:31285176-31285296;chrX:31285295-31285415;chrX:31285413-31285533;chrX:31285534-31285654;chrX:31285653-31285773;chrX:31285772-31285892;chrX:31285892-31286012;chrX:31286010-31286130;chrX:31286129-31286249;chrX:31286247-31286367;chrX:31286368-31286488;chrX:31286487-31286607;chrX:67543316-67543436;chrX:67543432-67543552;chrX:67543549-67543669;chrX:67543665-67543785;chrX:67543786-67543906;chrX:67543902-67544022;chrX:67544019-67544139;chrX:67544135-67544255;chrX:67544256-67544376;chrX:67544372-67544492;chrX:67544489-67544609;chrX:67544605-67544725;chrX:67544726-67544846;chrX:67544842-67544962;chrX:67544959-67545079;chrX:67545075-67545195;chrX:67545196-67545316;chrX:67545419-67545539;chrX:67545535-67545655;chrX:67545652-67545772;chrX:67545768-67545888;chrX:67545889-67546009;chrX:67546005-67546125;chrX:67546122-67546242;chrX:67546238-67546358;chrX:67546359-67546479;chrX:67546475-67546595;chrX:67546592-67546712;chrX:67546708-67546828;chrX:67546829-67546949;chrX:67546945-67547065;chrX:67547062-67547182;chrX:67547178-67547298;chrX:67547299-67547419;chrX:71449479-71449599;chrX:71449595-71449715;chrX:71450535-71450655;chrX:71450652-71450772;chrX:71450768-71450888;chrX:71450889-71451009;chrX:71451005-71451125;chrX:71451122-71451242;chrX:71451238-71451358;chrX:71451359-71451479;chrX:71453833-71453953;chrX:71453949-71454069;chrX:71454070-7145419; 0;chrX:71454186-71454306;chrX:71454419-71454539;chrX:71454540-71454660;chrX:71454656-71454776;chrX:71454773-71454893;chrX:71454889-71455009;chrX:71455010-71455130;chrX:137564825-137564945;chrX:137564943-137565063;chrX:137565060-137565180;chrX:137565178-137565298;chrX:137565299-137565419;chrX:137565416-137565536;chrX:137565534-137565654;chrX:137565651-137565771;chrX:137565772-137565892;chrX:137565890-137566010;chrX:137566007-137566127;chrX:137566125-137566245;chrX:137566246-137566366;chrX:137566363-137566483;chrX:137566481-137566601;chrX:137566598-137566718;chrX:137566719-137566839;chrX:137566837-137566957;chrX:137566954-137567074;chrX:137567072-137567192;chrX:137567193-137567313;chrX:137567310-137567430;chrX:137567428-137567548;chrX:137567545-137567665;chrX:137567666-137567786;chrX:137567784-137567904;chrX:137567901-137568021;chrX:137568019-137568139;chrX:137568140-137568260;chrX:137568258-137568378;chrX:137568375-137568495;chrX:137568493-137568613;chrX:137568614-137568734;chrX:137568732-137568852;chrX:140502316-140502436;chrX:140502432-140502552;chrX:140502549-140502669;chrX:140502665-140502785;chrX:140502786-140502906;chrX:140502902-140503022;chrX:140503019-140503139;chrX:140503135-140503255;chrX:140503256-140503376;chrX:140503372-140503492;chrX:140503489-140503609;chrX:140503605-140503725;chrX:140503726-140503846;chrX:140503842-140503962;chrX:140503959-140504079;chrX:140504075-140504195;chrX:140504196-140504316;chrX:140504381-140504501;chrX:140504497-140504617;chrX:140504614-140504734;chrX:140504730-140504850;chrX:140504851-140504971;chrX:140504967-140505087;chrX:140505084-140505204;chrX:140505200-140505320;chrX:140505321-140505441;chrX:140505437-140505557;chrX:140505554-140505674;chrX:140505670-140505790;chrX:140505791-140505911;chrX:140505907-140506027;chrX:140506024-140506144;chrX:140506140-140506260;chrX:140506261-140506381;chrX:147909978-; 147910098;chrX:147910094-147910214;chrX:147910211-147910331;chrX:147910327-147910447;chrX:147910448-147910568;chrX:147910564-147910684;chrX:147910681-147910801;chrX:147910797-147910917;chrX:147910918-147911038;chrX:147911034-147911154;chrX:147911151-147911271;chrX:147911267-147911387;chrX:147911388-147911508;chrX:147911504-147911624;chrX:147911621-147911741;chrX:147911737-147911857;chrX:147911858-147911978;chrX:147912111-147912231;chrX:147912227-147912347;chrX:147912344-147912464;chrX:147912460-147912580;chrX:147912581-147912701;chrX:147912697-147912817;chrX:147912814-147912934;chrX:147912930-147913050;chrX:147913051-147913171;chrX:147913167-147913287;chrX:147913284-147913404;chrX:147913400-147913520;chrX:147913521-147913641;chrX:147913637-147913757;chrX:147913754-147913874;chrX:147913870-147913990;chrX:147913991-147914111;chrX:148498601-148498721;chrX:148498717-148498837;chrX:148498834-148498954;chrX:148498950-148499070;chrX:148499071-148499191;chrX:148499187-148499307;chrX:148499304-148499424;chrX:148499420-148499540;chrX:148499541-148499661;chrX:148499657-148499777;chrX:148499774-148499894;chrX:148499890-148500010;chrX:148500011-148500131;chrX:148500127-148500247;chrX:148500244-148500364;chrX:148500360-148500480;chrX:148500481-148500601;chrX:148500743-148500863;chrX:148500859-148500979;chrX:148500976-148501096;chrX:148501092-148501212;chrX:148501213-148501333;chrX:148501329-148501449;chrX:148501446-148501566;chrX:148501562-148501682;chrX:148501683-148501803;chrX:148501799-148501919;chrX:148501916-148502036;chrX:148502032-148502152;chrX:148502153-148502273;chrX:148502269-148502389;chrX:148502386-148502506;chrX:148502502-148502622;chrX:148502623-148502743;chr13:102159577-102159697;chr13:102159695-102159815;chr13:102159812-102159932;chr13:102159930-102160050;chr13:102160051-102160171;chr13:102160169-102160289;chr13:102160286-102160406;chr13:10216040; <h2 style=";text-align:left;direction:ltr">4-102160524;chr13:102160525-102160645;chr13:102160643-102160763;chr13:102160760-102160880;chr13:102160878-102160998;chr13:102 160999-102161119;chr13:102161117-102161237;chr13:102161234-102 161354;chr13:102161352-102161472;chr13:102161473-102161593;chr1 3:102161708-102161828;chr13:102161826-102161946;chr13:102161947-102162067;chr13:102162065-102162185;chr13:102162182-102162302 ;chr13:102162300-102162420;chr13:102162421-102162541;chr13:102162539-102162659;chr13:102162656-102162776;chr13:102162774-1021 62894;chr13:102162895-102163015;chr13:102163013-102163133;chr13:102163130-102163250;chr13:102163248-102163368;chr13:102163369 -102163489;chr13:102163488-102163608;chr13:102163606-102163726;chr15:89331580-89331700;chr15:89331698-89331818;chr15:89331817 -89331937;chr15:89331935-89332055;chr15:89332056-89332176;chr1 5:89332175-89332295;chr15:89332295-89332415;chr15:89332413-8933 2533;chr15:89332532-89332652;chr15:89332650-89332770;chr15:893 32771-89332891;chr15:89332889-89333009;chr15:89333008-89333128;chr15:89333126-89333246;chr15:89333247-89333367;chr15:89333366-89333486;chr15:89333485-89333605;chr15:89333605-89 333725;chr15:89333723-89333843;chr15:89333842-89333962;ch r15:89333960-89334080;chr15:89334081-89334201;chr15:89334 199-89334319;chr15:89334318-89334438;chr15:89334436-89334 556;chr15:89334557-89334677;chr15:89334676-89334796;chr15 :89334795-89334915;chr15:89334915-89335035;chr15:89335033 -89335153;chr15:89335152-89335272;chr15:89335270-89335390 chr15:89335391-89335511 chr15:89335510-89335630 chr16:67841223-67841343 chr16:67841343-67841463 chr16:67841463-67841583 chr16:67841584-67841704 chr16:67841704-67841824 chr16:67841824-67841944 chr16:67841944-67842064 chr16:67842 065-67842185;chr16:67842185-67842305;chr16:67842305-67842425;chr16:67842426-67842546;chr16:67842546-67842666;chr16:67842666-67842786;chr16:67842786-67842906;chr16:67842907-67843027;chr16:67843027-67843147;chr16:67843147-6784326; 7;chr16:67843267-67843387;chr16:67843388-67843508;chr16:67843508-67843628;chr16:67843628-67843748;chr16:67843749-67843869;chr16:67843869-67843989;chr16:67843989-67844109;chr16:67844109-67844229;chr16:67844230-67844350;chr16:67844350-67844470;chr16:67844470-67844590;chr16:67844590-67844710;chr16:67844711-67844831;chr12:123531838-123531958;chr12:123531956-123532076;chr12:123532074-123532194;chr12:123532195-123532315;chr12:123532313-123532433;chr12:123532430-123532550;chr12:123532548-123532668;chr12:123532669-123532789;chr12:123532787-123532907;chr12:123532905-123533025;chr12:123533023-123533143;chr12:123533144-123533264;chr12:123533262-123533382;chr12:123533379-123533499;chr12:123533497-123533617;chr12:123533618-123533738;chr12:123533736-123533856;chr12:123533854-123533974;chr12:123533972-123534092;chr12:123534093-123534213;chr12:123534211-123534331;chr12:123534328-123534448;chr12:123534446-123534566;chr12:123534567-123534687;chr12:123534685-123534805;chr19:4510446-4510566;<h2 style=";text-align:left;direction:ltr">chr19:4510561-4510681;chr19:4510675-4510795;chr19:4510790-4510910;chr19:4510911-4511031;chr19:4511026-4511146;chr19:4511140-4511260;chr19:4511255-4511375;chr19:4511376-4511496;chr19:451149 1-4511611;chr19:4511605-4511725;chr19:4511720-4511840;chr19:4511841-4511961;chr19:4511956-4512076;chr19:4512070-4512190;chr19:4512185-4512305;chr19:4512306-4512426;chr19:4512420-4512540;chr 19:4512535-4512655;chr19:4512649-4512769;chr19:4512770-4512890;chr19:4512885-4513005;chr19:4512999-4513119;chr19:4513114-4513234;chr19:4513235-4513355;chr19:4513352-4513472;chr19:4513468-45 13588;chr19:4513584-4513704;chr7:55884919-55885039;chr7:55885040-55885160;chr7:55889640-55889760;chr7:55889746-55889866;chr7:55889853-55889973;chr7:55889959-55890079;chr7:55890520-55890640; 2) The 2× tiling probe design region includes the following gene loci: STR gene loci: TBP, FXN, NUTM2B-AS1 / LOC642361, NIPA1, SPAST, CSTB, YEATS2, NOTCH2NLC, ZNF713, PRDM12, EIF4A3; The specific sequences of 2× tiling probes are (a total of 217): chr1:149388722-149388842;chr1:149388782-149388902;chr1:149388842-149388962;chr1:149388902-149389022;chr1:149388962-149389082;chr1:149389022-149389142;chr1:149389082-149389202;chr1:149389142-149389262;chr1:149389202-149389322;chr1:149389262-149389382;chr1:149389322-149389442;chr1:149389382-149389502;chr1:149389442-149389562;chr1:149389502-149389622;chr1:149389562-149389682;chr1:149389622-149389742;chr1:149389682-149389802;chr1:149389742-149389862;chr1:149389802-149389922;chr1:149389862-149389982;chr1:149390762-149390882;chr10:79824234-79824354;chr10:79824294-79824414;chr10:79824354-79824474;chr10:79824414-79824534;chr10:79824474-79824594;chr10:79824534-79824654;chr10:79824594-79824714;chr10:79824654-79824774;chr10:79824714-79824834;chr10:79824774-79824894;chr10:79825014-79825134;chr10:79825074-79825194;chr10:79825134-79825254;chr10:79825194-79825314;chr10:79825254-79825374;chr10:79825314-79825434;chr10:79825374-79825494;chr10:79825434-79825554;chr10:79825494-79825614;chr10:79827344-79827464;chr10:79827404-79827524;chr10:79827464-79827584;chr10:79827524-79827644;chr10:79827584-79827704;chr10:79827644-79827764;chr10:79827704-79827824;chr10:79828064-79828184;chr10:79828124-79828244;chr10:79828184-79828304;chr10:79828244-79828364;chr10:79828304-79828424;chr10:79828364-79828484;chr15:22784771-22784891;chr15:22785491-22785611;chr15:22785551-22785671;chr15:22785611-22785731;chr15:22786091-22786211;chr15:22786151-22786271;chr15:22786211-22786331;chr15:22786271-22786391;chr15:22786331-22786451;chr15:22786391-22786511;chr15:22786451-22786571;chr15:22786511-22786631;chr15:22786571-22786691;chr15:22786631-22786751;chr17:80147039-80147159;chr17:80147099-80147219;chr17:80147159-80147279;chr17:80147219-80147339;chr17:80147279-80147399;chr17:80147339-80147459;chr17:80147399-80147519;chr17:80147459-80147579;chr17:80147519-80147639; chr17:80147579-80147699;chr17:80147639-80147759;chr17:80147699-80147819;chr17:80147999-80148119;chr17:80148059-80148179;chr17:80148119-80148239;chr17:80148359-80148479;chr17:80148419-80148539;chr2:32071717-32071837;chr2:32071777-32071897;chr2:32071837-32071957;chr2:32072317-32072437;chr2:32072377-32072497;chr2:32072557-32072677;chr2:32072617-32072737;chr2:32072677-32072797;chr2:32072737-32072857;chr2:32072797-32072917;chr2:32072857-32072977;chr2:32072917-32073037;chr2:32072977-32073097;chr2:32073037-32073157;chr2:32073097-32073217;chr2:32073277-32073397;chr2:32073337-32073457;chr2:32073397-32073517;chr2:32073577-32073697;chr2:32073637-32073757;chr2:32073697-32073817;chr2:32073757-32073877;chr2:32073817-32073937;chr2:32073877-32073997;chr2:32073937-32074057;chr2:32073997-32074117;chr2:32074057-32074177;chr2:32074117-32074237;chr2:32074177-32074297;chr2:32074237-32074357;chr2:32074297-32074417;chr2:32074357-32074477;chr2:32074417-32074537;chr2:32074777-32074897;chr2:32074837-32074957;<h2 style=";text-align:left;direction:ltr">chr2:32074897-32075017;chr2:32074957-32075077;chr2:320750 17-32075137;chr21:43776399-43776519;chr21:43776459-437765 79;chr21:43776519-43776639;chr21:43776579-43776699;chr21: 43776639-43776759;chr21:43776699-43776819;chr21:43776759-4 3776879;chr21:43776819-43776939;chr21:43776879-43776999;c hr21:43777959-43778079;chr21:43778019-43778139;chr21:43778 079-43778199;chr21:43778139-43778259;chr21:43778199-43778 319;chr21:43778259-43778379;chr21:43778319-43778439;chr21: 43778379-43778499;chr21:43778439-43778559;chr3:183712446- 183712566;chr3:183712506-183712626;chr3:183712566-18371268 6;chr3:183712626-183712746;chr3:183712686-183712806;chr3: 183713046-183713166;chr3:183713106-183713226;chr3:18371316 6-183713286;chr3:183713526-183713646;chr3:183713586-183713706;chr3:183713646-183713766;chr3:183713706-183713826;chr6:170559826-170559946;chr6:170559886-170560006;chr6:170559946-170560066;chr6:170560006-170560126;chr6:170560966-17; <h2 style=";text-align:left;direction:ltr">0561086;chr6:170561266-170561386;chr6:170561326-170561446;chr6 :170561386-170561506;chr6:170561446-170561566;chr6:170561506-1 70561626;chr6:170561566-170561686;chr6:170561626-170561746;chr 6:170561686-170561806;chr6:170561746-170561866;chr6:170561806- 170561926;chr7:55885581-55885701;chr7:55885641-55885761;chr7:55885701-55885821;chr7:55885761-55885881;chr7:55885821-55885941 ;chr7:55885881-55886001;chr7:55885941-55886061;chr7:55887081-55887201;chr7:55887141-55887261;chr7:55887201-55887321;chr7:5588 7261-55887381;chr7:55887321-55887441;chr7:55887381-55887501;chr7:55887441-55887561;chr7:55887501-55887621;chr9:69037474-6903 7594;chr9:69037534-69037654;chr9:69037594-69037714;chr9:69037894-69038014;chr9:69037954-69038074;chr9:69038014-69038134;chr9 :69038074-69038194;chr9:69038134-69038254;chr9:69038194-69038314;chr9:69038254-69038374;chr9:69038314-69038434;chr9:69038374 -69038494;chr9:69038434-69038554;chr9:69038734-69038854;chr9:69038794-69038914;chr9:69038854-69038974;chr9:69039214-69039334;<h2 style=";text-align:left;direction:ltr">chr9:69039274-69039394;chr9:130679945-130680065;chr9:130680665-130680785;chr9:130680725-130680845;chr9:130680785-130680905;chr9:130680845-130680965;chr9:130680905-130681025;chr9:130680965-130681085;chr9:130681025-130681145;c hr9:130681085-130681205;chr9:130681145-130681265;chr9:130681205-130681325;chr9:130681265-130681385;chr9:130681325-130681445;chr9:130681385-130681505;chr9:130681445-130681565;chr9:130681505-130681625;chr9:130681565-130681685; 3) The 3× tiling probe design region includes the following gene loci: STR gene loci: ATXN3, STARD7, DIP2B; VNTR gene locus: PLIN4; The specific sequences of 3× tiling probes are (a total of 109): chr12:50502921-50503041; chr12:50502961-50503081; chr12:50503001-50503121; chr12:50503041-50503161; c hr12:50503081-50503201;chr12:50503761-50503881;chr12:50503801-50503921;chr12:50503841-50503961;ch r12:50503881-50504001;chr12:50504521-50504641;chr12:50504561- 50504681;chr12:50504601-50504721;chr12:50504641-50504761;chr1 2:50504681-50504801;chr12:50504721-50504841;chr12:50504761-50 504881;chr12:50504801-50504921;chr12:50504841-50504961;chr12:5 0504881-50505001;chr12:50504921-50505041;chr12:50504961-50505 081;chr14:92068930-92069050;chr14:92068970-92069090;chr14:9206 9010-92069130;chr14:92070050-92070170;chr14:92070090-92070210 ;chr14:92070130-92070250;chr14:92070170-92070290;chr14:9207021 0-92070330;chr14:92070570-92070690;chr14:92070610-92070730;chr14:92070730-92070850;chr14:92070770-92070890;chr14:92070810-92070930;chr14:92070850-92070970;chr14:92070890-92071010;chr14:92070930-92071050;chr14:92070970-92071090;chr19:4508527-4508 647;chr19:4508567-4508687;chr19:4508607-4508727;chr19:4508647 -4508767;chr19:4508687-4508807;chr19:4508727-4508847;chr19:450 8767-4508887;chr19:4508807-4508927;chr19:4508847-4508967;chr1 9:4508887-4509007;chr19:4508927-4509047;chr19:4508967-4509087;<h2 style=";text-align:left;direction:ltr">chr19:4510327-4510447;chr19:4510367-4510487;chr19:4510407-4510527;chr19:4510447-4510567;chr19:4513501-4513621;chr19:4513541-4513661;chr19:4513581-4513701;chr19:4513621-4513741;chr19:4513661-4513781;chr19:4513701-4513821;chr19:4513741 -4513861;chr19:4513781-4513901;chr19:4513821-4513941;chr19:4513861-4513981;chr19:4513901-4514021;chr19:4513941-4514061;chr19:4513981-4514101;chr19:4514021-4514141;chr19:4514061-4514181;chr19:4514101-4514221;chr19:4515541-4515661;chr2: 96194975-96195095;chr2:96195015-96195135;chr2:96195055-961 95175;chr2:96195095-96195215;chr2:96195135-96195255;chr2:9 6195175-96195295;chr2:96195215-96195335;chr2:96195255-9619 5375;chr2:96195295-96195415;chr2:96195335-96195455;chr2:961 95375-96195495;chr2:96195415-96195535;chr2:96195455-961955 75;chr2:96195495-96195615;chr2:96195535-96195655;chr2:96195 575-96195695;chr2:96195615-96195735;chr2:96195655-96195775 ;chr2:96195695-96195815;chr2:96195735-96195855;chr2:961974; 15-96197535;chr2:96197455-96197575;chr2:96197495-96197615;chr2:96197535-96197655;chr2:96197575-96197 695;chr2:96197935-96198055;chr2:96197975-96198095;chr2:96198375-96198495;chr2:96198415-96198535;chr2: 96198455-96198575;chr2:96198495-96198615;chr2:96198775-96198895;chr2:96198815-96198935;chr2:96198855- 96198975;chr2:96198895-96199015;chr2:96198935-96199055;chr2:96199055-96199175;chr2:96199095-96199215; 4) The 4× tiling probe design region includes the following gene loci: STR gene loci: ATXN10, ZNF713; The specific sequences of 4× tiled probes are (37 in total): chr22:45795329-45795449; chr22:45796769-45796889; chr22:45796799-45796919; chr22:45796829-45796949; chr22:457968 59-45796979;chr22:45796889-45797009;chr22:45796919-45797039;chr22:45796949-45797069;chr22:45796979-45797099;c hr22:45797009-45797129; chr22:45797039-45797159; chr22:45797069-45797189; chr22:45797099-45797219; chr22:4579712 9-45797249;chr22:45797159-45797279;chr22:45797189-45797309;chr22:45797219-45797339;chr7:55887874-55887994;chr 7:55887904-55888024;chr7:55887934-55888054;chr7:55887964-55888084;chr7:55887994-55888114;chr7:55888024-55888 144;chr7:55888054-55888174;chr7:55888084-55888204;chr7:55888114-55888234;chr7:55888204-55888324;chr7:55888234 -55888354;chr7:55888264-55888384;chr7:55888294-55888414;chr7:55889434-55889554;chr7:55889464-55889584;chr7:5 5889494-55889614;chr7:55889524-55889644;chr7:55889554-55889674;chr7:55889584-55889704;chr7:55889614-55889734.

6. The probe set according to any one of claims 0-5, wherein each probe in the probe set is at least one of a double-stranded DNA, a single-stranded DNA or a single-stranded RNA; wherein both the forward sequence and the reverse complementary sequence of the double-stranded DNA probe can be used to capture dynamically mutated STR and VNTR gene sites.

7. The probe set according to any one of claims 0 to 6, wherein each probe in the probe set carries a label.

8. The probe set according to any one of claims 0 to 7, wherein the label is selected from one or more of the following groups: biotin, avidin, antibiotin protein, antibody or chemical coupling agent; preferably biotin.

9. A method for constructing a single-molecule long-read sequencing library, comprising the following steps: 1) Randomly fragment the DNA sample; 2) performing end repair on the fragmented DNA and ligating the pre-library adapter to obtain a pre-library; 3) hybridizing the prelibrary with the probe set in the absence of a blocking sequence to obtain a hybridization product; 4) Using magnetic beads to capture the hybridization products and after washing, PCR amplification is performed to obtain a capture library; 5) Construction of third-generation sequencing library.

10. The method according to claim 9, wherein the random DNA fragmentation method in step 1) comprises ultrasonic treatment, mechanical disruption or enzyme cleavage.

11. The method according to claim 9, wherein the randomly fragmented DNA in step 1) has a length ranging from 5 to 15 kb.

12. The method according to claim 11, wherein the length of the randomly fragmented DNA in step 1) is preferably in the range of 7-10 kb.

13. The method according to claim 9, wherein the pre-library adapter in step 2) is in a Y-shaped structure, which can be an overhanging T adapter or a blunt-end adapter.

14. The method according to claim 13, wherein the pre-library adapter comprises two incompletely complementary DNA single strands, the length of the DNA single strands may be 30-70 bp, and is prepared into a working adapter after annealing reaction.

15. The method according to claim 9, wherein in step 2), the pre-library can be obtained without performing pre-capture PCR amplification.

16. The method according to claim 9, wherein the probe group in step 3) is the probe group according to any one of claims 0-8.

17. The method according to claim 9, wherein the hybridization system in step 3) comprises a hybridization buffer, an enhancement solution, human Cot-1 DNA and the probe set according to any one of claims 0 to 8, but does not include a blocking sequence.

18. The method according to claim 9, wherein the blocking sequence in step 3) is a sequence designed to be reverse complementary to the adapter sequence.

19. The method according to claim 9, wherein the amplification system in step 4) comprises an amplification enzyme, an amplification buffer, a PCR additive, an amplification primer and a magnetic bead to capture the product.

20. The method according to claim 19, wherein the amplification enzyme has high fidelity and excellent amplification properties, and is suitable for amplifying complex templates such as long fragments and high GC / AT content.

21. The method according to claim 19, wherein the PCR additive can reduce secondary structure, enhance enzyme stability or inhibit non-specific amplification; the PCR additive is selected from at least one of dimethyl sulfoxide, betaine, bovine serum albumin and formamide.

22. The method according to claim 19, wherein the amplification primer has a final concentration range of 0.1-1 μM, is phosphorylated, and is compatible with the pre-library adapter; the amplification procedure comprises: 94℃ 2-4min; 98℃ 10-15sec, Tm℃ 30sec, 68℃ 5-10min, 15-25cycles; 68℃ 5-10min.

23. The method according to claim 9, wherein the main band of the third-generation sequencing library in step 5) is about 8 kb.

24. A device for detecting the number of repetitions of STR and VNTR gene loci, comprising: 1) Capture module: used to construct a capture library; 2) Library construction module: used to construct single-molecule long-read sequencing libraries; 3) Sequencing and analysis module: perform third-generation sequencing on the sequencing library and perform data analysis to obtain the sequence information of the target repetitive region.

25. The device according to claim 24, wherein the probe set used to construct the capture library is the probe set according to any one of claims 0-8.

26. The device according to claim 24, wherein the method for constructing a capture library and a single-molecule long-read sequencing library is the method according to any one of claims 9 to 23.

27. The device according to claim 24, wherein the single-molecule long-read sequencing platform is selected from the Pacific Biosciences SMRT sequencing platform based on single-molecule real-time sequencing technology and the Oxford Nanopore single-molecule sequencing platform.

28. The device according to claim 24, wherein the data analysis process comprises: 1) After the third-generation sequencing data is split into samples, it is compared with the human genome reference sequence; 2) retain the uniquely aligned sequences and screen the target regions; 3) Count the number of target repeat region sequences, capture efficiency and coverage depth.

29. According to the process of claim 28, the method for detecting the number of repetitions of STR gene loci is: 1) Based on the sequence of the target region, analyze the repeat region, calculate the number of repeats and supporting sequences, and determine the pathogenicity based on the information included in the database; 2) Based on the specific sequence alignment of the 5' flanking region and the 3' flanking region of the target repeat region, the repeat region sequence is cut, the number of repeat units and the typing clustering are analyzed, the repeat structure of each allele is obtained, and the number of repeats of each allele and the number of sequences supporting the allele are counted.

30. The method according to claim 29, wherein the method for detecting the number of repetitions of the STR gene loci further comprises: when the number of repetitions of two alleles of the STR gene locus is the same, phasing is performed according to the SNVs of the flanking regions on both sides of the repetition region to split the haplotype; for two alleles with the same repetition structure, the number of repetitions and the number of supporting sequences are counted respectively.

31. The method according to any one of claims 29 or 30, wherein the method for detecting the number of repetitions of the STR gene loci further comprises, when the STR gene loci are positively highly repeated and interrupted, aligning the flanks of the repeat region of the unilateral repeat sequence, locating the upstream or downstream of the repeat region, cutting and filtering, calculating the number of repeat units, and supplementing the pathogenicity determination.

32. According to the process of claim 28, the method for detecting the number of repetitions of the VNTR gene locus is: 1) Construct a list of repeat types for each VNTR gene based on the existing sample set; 2) Based on the specific sequence alignment of the 5' flanking region and the 3' flanking region of the target repeat region, the repeat region sequence is cut, the repeat unit is identified from the built-in VNTR gene repeat type list, and the same type of polymorphisms are combined to calculate the number of repeats; 3) Typing and clustering, counting the number of repetitions and supporting sequences of each allele.

33. A method for detecting the number of repetitions of dynamic mutation STR and VNTR gene loci, comprising the following steps: 1) Perform random fragmentation, end repair and pre-library adapter ligation on DNA samples to obtain Get the pre-library; 2) in the absence of a blocking sequence, hybridizing the probe set according to any one of claims 1 to 8 with the prelibrary, capturing the hybridization product with magnetic beads, and performing PCR amplification after washing to obtain a captured library; 3) Construct a third-generation sequencing library, perform third-generation sequencing and analyze the sequencing data to obtain the target sequence information.

34. The method according to claim 33, wherein the method for constructing the pre-library, the capture library and the third-generation sequencing library is the method according to any one of claims 9 to 23.

35. The method according to claim 33, wherein the sequencing data analysis method is the method according to any one of claims 28 to 32.

36. A kit for detecting dynamic mutation STR and VNTR gene loci, comprising: 1) The probe set according to any one of claims 0 to 8; 2) Reagents for constructing a prelibrary; 3) Reagents for hybridization capture, excluding blocking sequences; 4) Reagents for PCR amplification; 5) Reagents for constructing third-generation sequencing libraries.

37. The kit according to claim 36, wherein the reagents for constructing the pre-library include a repair enzyme, a repair buffer, a ligase, a ligation buffer and a pre-library adapter.

38. The kit according to claim 37, wherein the pre-library adapter is in a Y-shaped structure, which can be an overhanging T adapter or a blunt-end adapter.

39. The kit according to claim 36, wherein the hybridization capture reagent comprises a hybridization buffer, an enhancement solution, human Cot-1 DNA, the probe set according to any one of claims 0-8, magnetic beads and a washing reagent, but does not comprise a blocking sequence.

40. The kit according to claim 39, wherein the blocking sequence is a sequence designed to be reverse complementary to the linker sequence.

41. The kit according to claim 36, wherein the PCR amplification reagents include an amplification enzyme, an amplification buffer, a PCR additive and an amplification primer.

42. The kit according to claim 41, wherein the amplification enzyme has high fidelity and excellent amplification properties, and is suitable for amplifying complex templates such as long fragments and high GC / AT content.

43. The kit according to claim 41, wherein the PCR additive can reduce secondary structure, enhance enzyme stability or inhibit nonspecific amplification, and the PCR additive is selected from at least one of dimethyl sulfoxide, betaine, bovine serum albumin, and formamide.

44. The kit according to claim 41, wherein the amplification primer has a final concentration range of 0.1-1 μM, is phosphorylated and is compatible with the pre-library adapter.

45. The kit according to claim 36, wherein the reagents for constructing a third-generation sequencing library include a repair enzyme, a ligase, an exonuclease and a matching buffer, and a third-generation sequencing adapter.

46. ​​Use of the probe set described in any one of claims 0-8, or the single-molecule long-read sequencing library construction method described in any one of claims 9-23, or the device described in any one of claims 24-32, or the method described in any one of claims 33-35 in the preparation of a kit for detecting dynamic mutation STR and VNTR gene sites.

47. According to the use of claim 46, the STR and VNTR gene loci include: 1) STR gene loci (69): ATXN1, ATXN2, ATXN3, CACNA1A, ATXN7, ATXN8OS / ATXN8, ATXN10, PPP2R2B, TBP, BEAN1 / TK2, NOP56, DAB1, ATN1, FXN, GLS, RFC1, AR, DMPK, CNBP, LRP12, GIPC1, NUTM2B-AS1 / LOC642361, PABPN1, C9orf72, NIPA1, TAF1, DMD, VWA1, SPAST, CSTB, SAMD1 2. STARD7, MARCHF6, YEATS2, TNRC6A, RAPGEF2, NOTCH2NLC, FMR1, AFF2, AFF3, ZNF713, C11ORF80, DIP2B, CBL, HTT, JPH3, PRDM12, PR NP, SOX3, ARX, ZIC2, HOXD13, EIF4A3, RUNX2, HOXA13, FOXL2, ZIC3, TBX1, XYLT1, COMP, TCF4, PHOX2B, FGF14, RILPL1, THAP11, POLG; 2) VNTR gene loci (4): CACNA1C, PLIN4, ABCA7, and WDR7.

Images