CN101921842B - Hla-a,b基因分型用pcr引物及其使用方法 - Google Patents
Hla-a,b基因分型用pcr引物及其使用方法 Download PDFInfo
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- CN101921842B CN101921842B CN 201010213721 CN201010213721A CN101921842B CN 101921842 B CN101921842 B CN 101921842B CN 201010213721 CN201010213721 CN 201010213721 CN 201010213721 A CN201010213721 A CN 201010213721A CN 101921842 B CN101921842 B CN 101921842B
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- C12Q1/6881—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
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Abstract
Description
引物标签 编号 | 正向引物标签 | 反向引物标签 | 对应96孔板 位置 | 对应模板 (组1) |
PI-1 | TCGCAGACATCA | TGACACGATGCT | A1 | 1 |
PI-2 | TACATCGCACTA | TACAGATGCTGA | A2 | 2 |
PI-3 | CTCGATGAGTAC | ACGTCTAGACAC | A3 | 3 |
PI-4 | TCTGTATACTCA | TGCTGTAGTGAC | A4 | 4 |
PI-5 | TATCTGCTCATA | AGATATCGAGCT | A5 | 5 |
PI-6 | TACATGCTGAGC | ACGTGTCTATCA | A6 | 6 |
PI-7 | TCATATCGCGAT | AGATCGTATAGC | A7 | 7 |
PI-8 | ACAGATGCACGC | ATCTCGTGACAG | A8 | 8 |
PI-9 | TAGATCGTACAT | ACTAGTACACGC | A9 | 9 |
PI-10 | ACTACACGTCTC | ATAGTCACGCGT | A10 | 10 |
PI-11 | AGACTCGCGTAT | TACTAGCTGACG | A11 | 11 |
PI-12 | ATACTAGTGCTC | TGTATCGTGCTC | A12 | 12 |
PI-13 | CACGATGACATC | TAGTGAGCGCAC | B1 | 13 |
PI-14 | TGCTGTCTCGAG | CATAGCAGTGTC | B2 | 14 |
PI-15 | TGTGCTCGAGTC | TCTGATCGAGCA | B3 | 15 |
PI-16 | CACTCGTACATC | AGCGATGCTCAT | B4 | 16 |
PI-17 | CGACGTGCTCGC | CGCGTACTGCAG | B5 | 17 |
PI-18 | ACGCATCTATAC | CTAGTATCGCAG | B6 | 18 |
PI-19 | CGAGATGACTCT | TGTATACACGAT | B7 | 19 |
PI-20 | ACTGTCTCGAGC | ACGTAGCGCACA | B8 | 20 |
PI-21 | CATCTGCTATAG | TCTAGCTCATGA | B9 | 21 |
PI-22 | ACGCACTCTAGA | CTATGCACTGAT | B10 | 22 |
PI-23 | TGAGATACAGTA | ATCTGCTATGAC | B11 | 23 |
PI-24 | ACTCATCGTGCT | TAGAGCTGTCAC | B12 | 24 |
PI-25 | TACACTGTCTAT | CAGCACATAGAT | C1 | 25 |
PI-26 | CACAGTACTCGC | CTGCTAGTGTAT | C2 | 26 |
PI-27 | TGTACTATCATA | TGTGATAGACAC | C3 | 27 |
PI-28 | CTAGTACTGACG | AGCGAGTCTACT | C4 | 28 |
PI-29 | TAGACTGAGCTA | ACATACTGAGAC | C5 | 29 |
PI-30 | CAGACGCGTGAG | TACATCTCGTAT | C6 | 30 |
PI-31 | CGCGACATCACG | TAGCGATGAGAC | C7 | 31 |
PI-32 | ACACTCATAGAT | CTATCATGACAC | C8 | 32 |
PI-33 | AGCGTATACTAG | CATACTCACGTA | C9 | 33 |
PI-34 | TGTCGTGCTATC | ACATGACTCACG | C10 | 34 |
PI-35 | CGCTAGACTGTA | TACTATAGTCGA | C11 | 35 |
PI-36 | ACAGTGTAGCGC | TGATATGCTACA | C12 | 36 |
PI-37 | CACTCTATCGAC | TCACGCGATGAG | D1 | 37 |
PI-38 | ACACTCTAGTCA | ACGTAGATCTAT | D2 | 38 |
PI-39 | CATATGAGATCG | AGCAGAGTGCTC | D3 | 39 |
PI-40 | CAGCTATCATAC | CACTGCAGACGA | D4 | 40 |
PI-41 | TATACTCTAGAT | TGCATAGAGCGC | D5 | 41 |
PI-42 | TGTATGCTCGTC | TCGTGACAGATC | D6 | 42 |
PI-43 | TAGTGATGCTCT | ACGAGCTGATAT | D7 | 43 |
PI-44 | AGACTCTGAGTC | CTGATAGTATCA | D8 | 44 |
PI-45 | CTCATAGACTAC | ATCGCGAGTGAC | D9 | 45 |
PI-46 | TCGCTCACTACA | TGTCTCGACATC | D10 | 46 |
PI-47 | ATAGAGTCTCAT | CGCATAGCGTAT | D11 | 47 |
PI-48 | CGAGACACTCGC | TCGTAGTCTACA | D12 | 48 |
PI-49 | CAGCATACTATC | TCGTGATACAGA | E1 | 49 |
PI-50 | CAGCTATAGTCT | ATGCAGATATCT | E2 | 50 |
PI-51 | TCTATCGATGCA | ACACGCAGATCG | E3 | 51 |
PI-52 | CATGAGTATAGC | CTAGCTGACGTA | E4 | 52 |
PI-53 | TAGCATATCGAG | TACACGTATGAG | E5 | 53 |
PI-54 | ACGACTCGCTAC | TCATGACTAGTA | E6 | 54 |
PI-55 | TAGCATACACGC | TGACGCGTATAC | E7 | 55 |
PI-56 | CGTCATATGCAG | TATAGCGATGAC | E8 | 56 |
PI-57 | TGCAGCGAGTAC | TCGACGCTAGCG | E9 | 57 |
PI-58 | CGTGTCGACAGA | CAGTCGTGAGCA | E10 | 58 |
PI-59 | ACTCGACGTGAG | ACGCGAGTGATA | E11 | 59 |
PI-60 | ACTCGTCTGACG | TGCTATCACTGA | E12 | 60 |
PI-61 | CATACTGTATCT | TACATAGATGTC | F1 | 61 |
PI-62 | TCTACTCGTGAC | CACGTATAGTGA | F2 | 62 |
PI-63 | CTGCACTAGACA | ACTCATATCGCA | F3 | 63 |
PI-64 | ACACGAGCTCAT | CACTCATATCGA | F4 | 64 |
PI-65 | TACAGATAGTCT | TCGTCTGTGATA | F5 | 65 |
PI-66 | TACACTCGTGCT | TGACGCTCATCT | F6 | 66 |
PI-67 | TACATGTGACGA | TCGTACATGCTC | F7 | 67 |
PI-68 | TGTATGATCTCG | CACTGTGCTCAT | F8 | 68 |
PI-69 | CAGTACACTCTA | ACTGCATGATCG | F9 | 69 |
PI-70 | CATACTATCACG | TCGTGTCACTAC | F10 | 70 |
PI-71 | CACTATACAGAT | CGACACGTACTA | F11 | 71 |
PI-72 | ATATCGTAGCAT | TCGTGATCACTA | F12 | 72 |
PI-73 | TAGTCTATACAT | AGACGCTGTCGA | G1 | 73 |
PI-74 | TGTCACAGTGAC | TCATATGATCGA | G2 | 74 |
PI-75 | ATCGACTATGCT | CGATCATATGAG | G3 | 75 |
PI-76 | ATACTAGCATCA | TCATGCTGACGA | G4 | 76 |
PI-77 | CACTGACGCTCA | CACTACATCGCT | G5 | 77 |
PI-78 | TCGCTCATCTAT | TAGTACAGAGCT | G6 | 78 |
PI-79 | TGTATCACGAGC | ATGATCGTATAC | G7 | 79 |
PI-80 | TACTGCTATCTC | CGCTGCATAGCG | G8 | 80 |
PI-81 | CGCGAGCTCGTC | ACTCGATGAGCT | G9 | 81 |
PI-82 | TAGAGTCTGTAT | TGTCTATCACAT | G10 | 82 |
PI-83 | TACTATCGCTCT | TATGTGACATAC | G11 | 83 |
PI-84 | TAGATGACGCTC | TACTCGTAGCGC | G12 | 84 |
PI-85 | TCGCGTGACATC | ATCTACTGACGT | H1 | 85 |
PI-86 | ACACGCTCTACT | ACAGTAGCGCAC | H2 | 86 |
PI-87 | TACATAGTCTCG | CTAGTATCATGA | H3 | 87 |
PI-88 | TGAGTAGCACGC | TCGATCATGCAG | H4 | 88 |
PI-89 | TAGATGCTATAC | TACATGCACTCA | H5 | 89 |
PI-90 | ATCGATGTCACG | CAGCTCGACTAC | H6 | 90 |
PI-91 | ATCATATGTAGC | CTCTACAGTCAC | H7 | 91 |
PI-92 | TAGCATCGATAT | AGATAGCACATC | H8 | 92 |
PI-93 | TGATCGACGCTC | CTAGATATCGTC | H9 | 93 |
PI-94 | TGCAGCTCATAG | TACAGACTGCAC | H10 | 94 |
PI-95 | CGACGTAGAGTC | CAGTAGCACTAC | H11 | 95 |
Promega 5×buffer I(Mg2+plus) | 5.0ul |
dNTP Mixture(各2.5mM) | 2.0ul |
PInf-A/B-F2/3/4(2pmol/ul) | 1.0ul |
PInr-A/B-R2/3/4(2pmol/ul) | 1.0ul |
Promega Taq(5U/ul) | 0.2ul |
DNA(约20ng/ul) | 5.0ul |
ddH2O | 10.8ul |
Total | 25.0ul |
Promega 5×buffer I(Mg2+plus) | 5.0ul |
dNTP Mixture(各2.5mM) | 2.0ul |
PInf-D2-F1(2pmol/ul) | 1.0ul |
PInf-D2-F2(2pmol/ul) | 1.0ul |
PInf-D2-F3(2pmol/ul) | 1.0ul |
PInf-D2-F4(2pmol/ul) | 1.0ul |
PInf-D2-F5(2pmol/ul) | 1.0ul |
PInf-D2-F6(2pmol/ul) | 1.0ul |
PInf-D2-F7(2pmol/ul) | 1.0ul |
PInr-D2-R(2pmol/ul) | 1.0ul |
Promega Taq(5U/ul) | 0.2ul |
DNA(约20ng/ul) | 5.0ul |
ddH2O | 4.8ul |
Total | 25.0ul |
负载比(Duty Cycle) | 10% |
强度(Intensity) | 5 |
循环/脉冲(Cycles/Burst) | 200 |
时间(秒)(Time seconds) | 300 |
qPCR检测结果(nM) | |
HLA-Mix | 78.90 |
Claims (7)
Priority Applications (14)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 201010213721 CN101921842B (zh) | 2010-06-30 | 2010-06-30 | Hla-a,b基因分型用pcr引物及其使用方法 |
PCT/CN2010/001813 WO2012000150A1 (zh) | 2010-06-30 | 2010-11-12 | Hla-a,b基因分型用pcr引物及其使用方法 |
MYPI2012005591A MY173793A (en) | 2010-06-30 | 2011-06-30 | New pcr sequencing method and use thereof in hla genotyping |
PCT/CN2011/076688 WO2012000445A1 (zh) | 2010-06-30 | 2011-06-30 | 新的的pcr测序方法及其在hla基因分型中的应用 |
JP2013516983A JP5968879B2 (ja) | 2010-06-30 | 2011-06-30 | Dna分子タグ技術及びdna不完全断片化技術に基づいたpcrシークエンシング法及びそれを用いたhla遺伝子タイピング法 |
EP11800190.8A EP2599877B1 (en) | 2010-06-30 | 2011-06-30 | New pcr sequencing method and use thereof in hla genotyping |
DK11800190.8T DK2599877T3 (da) | 2010-06-30 | 2011-06-30 | Hidtil ukendt pcr sekvenseringsfremgangsmåde og anvendelse deraf i hla-genotypebestemmelse |
CA2803940A CA2803940C (en) | 2010-06-30 | 2011-06-30 | Application of a pcr sequencing method, based on dna barcoding technique and dna incomplete shearing strategy, in hla genotyping |
AU2011274090A AU2011274090B2 (en) | 2010-06-30 | 2011-06-30 | New PCR sequencing method and use thereof in HLA genotyping |
US13/807,660 US9957564B2 (en) | 2010-06-30 | 2011-06-30 | Application of a PCR sequencing method, based on DNA barcoding technique and DNA incomplete shearing strategy, in HLA genotyping |
BR112012032586-8A BR112012032586B1 (pt) | 2010-06-30 | 2011-06-30 | Métodos para determinar a sequência nucleotídica de um ácido nucléico de interesse e para determinar o genótipo hla em uma amostra |
RU2013103795/10A RU2587606C2 (ru) | 2010-06-30 | 2011-06-30 | Новый способ пцр-секвенирования и его применение в генотипировании hla |
SG2012096616A SG186876A1 (en) | 2010-06-30 | 2011-06-30 | New pcr sequencing method and use thereof in hla genotyping |
KR1020137002332A KR101709826B1 (ko) | 2010-06-30 | 2011-06-30 | 신규 pcr 서열화 방법 및 hla 제노타이핑에서의 상기 방법의 사용 |
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CN 201010213721 CN101921842B (zh) | 2010-06-30 | 2010-06-30 | Hla-a,b基因分型用pcr引物及其使用方法 |
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CN101921842B true CN101921842B (zh) | 2013-08-07 |
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WO2012083506A1 (zh) * | 2010-12-24 | 2012-06-28 | 深圳华大基因科技有限公司 | Hla-dqb1基因分型的方法及其相关引物 |
CN101921841B (zh) * | 2010-06-30 | 2014-03-12 | 深圳华大基因科技有限公司 | 基于Illumina GA测序技术的HLA基因高分辨率分型方法 |
CA2803940C (en) | 2010-06-30 | 2019-07-02 | Bgi Shenzhen Co., Limited | Application of a pcr sequencing method, based on dna barcoding technique and dna incomplete shearing strategy, in hla genotyping |
CN103261438B (zh) * | 2010-12-24 | 2015-09-16 | 深圳华大基因医学有限公司 | Hla-c基因分型的方法及其相关引物 |
CN110621785B (zh) * | 2017-06-20 | 2023-08-15 | 深圳华大生命科学研究院 | 基于三代捕获测序对二倍体基因组单倍体分型的方法和装置 |
CN107937487B (zh) * | 2017-12-29 | 2019-06-21 | 北京诺诗康瀛基因技术股份有限公司 | 一种用于hla-a基因pcr扩增、基因分型的方法、引物组及试剂盒 |
CN108441547B (zh) * | 2018-04-13 | 2020-02-07 | 北京诺诗康瀛基因技术股份有限公司 | 一种hla基因扩增、基因分型的引物组、试剂盒和方法 |
CN111235266B (zh) * | 2020-03-10 | 2023-12-01 | 广州医科大学附属第二医院 | Hla亚型检测试剂盒及其应用 |
CN112921078B (zh) * | 2021-02-08 | 2022-07-08 | 杭州虎贲生物医疗有限公司 | 一种测定待测者的hla-i分型的方法和一种锚序列组 |
CN113278687B (zh) * | 2021-05-20 | 2023-11-24 | 广州医科大学附属第二医院 | 用于hla-b1502和hla-a2402基因型检测的试剂盒 |
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CN1451760A (zh) * | 2002-04-16 | 2003-10-29 | 中国人民解放军军事医学科学院放射医学研究所 | Hla分型用寡核苷酸芯片的制备及用法 |
GB0400584D0 (en) * | 2004-01-12 | 2004-02-11 | Solexa Ltd | Nucleic acid chacterisation |
CA2701411A1 (en) * | 2007-10-16 | 2009-04-23 | F. Hoffmann-La Roche Ag | High resolution, high throughput hla genotyping by clonal sequencing |
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CN101313078A (zh) * | 2005-09-29 | 2008-11-26 | 科因股份有限公司 | 突变群体的高通量筛选 |
CN101654691A (zh) * | 2009-09-23 | 2010-02-24 | 深圳华大基因科技有限公司 | Hla基因扩增和基因分型方法及其相关引物 |
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