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    METHOD AND REAGENT FOR CONSTRUCTING NUCLEIC ACID DOUBLE-LINKER SINGLE-STRAND CYCLICAL LIBRARY
    2.
    发明申请
    METHOD AND REAGENT FOR CONSTRUCTING NUCLEIC ACID DOUBLE-LINKER SINGLE-STRAND CYCLICAL LIBRARY 审中-公开

    公开(公告)号：US20170349893A1

    公开(公告)日：2017-12-07

    申请号：US15529867

    申请日：2014-11-26

    申请人： BGI SHENZHEN ,  BGI SHENZHEN CO., LIMITED

    发明人： Yuan JIANG ,  Xia ZHAO ,  Andrei ALEXEEV ,  Radoje DRMANAC ,  Wenwei ZHANG ,  Hui JIANG

    IPC分类号： C12N15/10

    CPC分类号： C12N15/1068 ,  C12N15/10 ,  C12N15/11 ,  C12Q1/68 ,  C40B50/06 ,  C40B50/18

    摘要： A method and reagent for constructing a nucleic acid double-joint single-strand cyclical library. The method comprises: breaking a nucleic acid into nucleic acid fragments; connecting a first linker sequence; producing by amplification a first product provided with the first linker sequence at either end, where a U nucleobase is provided on a primer sequence; using USER enzyme to cleave the first product and cyclizing to produce a gap; or, a nicking enzyme recognition sequence is also provided on the primer sequence, using the USER enzyme to cleave the first product, cyclizing and using a nicking enzyme for nicking to produce a nick; performing a restrictive nick/gap translation reaction from the nick or the gap; removing by digestion any portion that did not undergo the restrictive nick/gap translation reaction; connecting a second linker sequence; producing by amplification a second product provided with the second linker sequence at either end; denaturing the second product, and using a mediated sequence for cyclization of a single-strand nucleic acid molecule. The method allows an increase in the length of library insert fragments and obviates the need for gel extraction; the single-strand nucleic acid molecule can be cyclized directly when denatured with heat.

    Method and reagent for constructing nucleic acid double-linker single-strand cyclical library
    4.
    发明授权
    Method and reagent for constructing nucleic acid double-linker single-strand cyclical library 有权

    公开(公告)号：US10479991B2

    公开(公告)日：2019-11-19

    申请号：US15529881

    申请日：2014-11-26

    申请人： BGI SHENZHEN ,  BGI SHENZHEN CO., LIMITED

    发明人： Yuan Jiang ,  Qiaoling Li ,  Andrei Alexeev ,  Evan Hurowitz ,  Xia Zhao ,  Tong Wang ,  Chao Dong ,  Dong Li ,  Radoje Drmanac ,  Wenwei Zhang ,  Hui Jiang

    IPC分类号： C12N15/10 ,  C40B50/18 ,  B01J19/00 ,  C40B50/06

    摘要： A method and reagent for constructing a nucleic acid double-linker single-strand cyclic library. The method comprises: breaking a nucleic acid into nucleic acid fragments; connecting a first linker sequence; producing by amplification a first product provided with the first linker sequence at either end, where a U nucleobase site is provided on primer sequences and a nicking enzyme recognition sequence is either provided or not provided on same, and a first affinity tag is provided on one of the primer sequences; using USER enzyme to cleave the first product; cyclizing the cleaved first product; treating the cyclization product with either a phosphatase or a nicking enzyme; using a solid-phase vector for combination with a cyclized molecule; performing a restrictive gap translation reaction; removing by digestion any portion that did not undergo the restrictive gap translation reaction; connecting a second linker sequence; producing by amplification a second product provided with the second linker sequence at either end; denaturing the second product, and cyclizing a single-strand nucleic acid molecule. The method allows an increase in the length of library insert fragments, a simplified library construction process, reduced library construction time, and reduced library construction costs.

    TARGET REGION ENRICHMENT METHOD BASED ON MULTIPLEX PCR, AND REAGENT
    5.
    发明申请
    TARGET REGION ENRICHMENT METHOD BASED ON MULTIPLEX PCR, AND REAGENT 审中-公开

    公开(公告)号：US20180163251A1

    公开(公告)日：2018-06-14

    申请号：US15537396

    申请日：2015-12-09

    申请人： BGI SHENZHEN ,  BGI SHENZHEN CO., LIMITED

    发明人： Jing Guo ,  Rongrong GUO ,  Meiyan LI ,  Chunyu GENG ,  Hui JIANG

    IPC分类号： C12Q1/6806 ,  C12P19/34

    CPC分类号： C12Q1/6806 ,  C12P19/34 ,  C12Q1/6855 ,  C12Q2525/155 ,  C12Q2531/101 ,  C12Q2563/131 ,  C12Q2563/179

    摘要： Provided are a target region enrichment method based on multiplex PCR, and a reagent, the method comprising: connecting a first linker and a second linker respectively at two ends of a nucleic acid segment containing target regions to be enriched so as to obtain a linker-connected product; performing a PCR amplification on the linker-connected product using a first primer specifically bound to the first linker and a second primer specifically bound to the second linker to obtain an amplified product, the first primer or the second primer having a first affinity label; capturing a single strand having the first affinity label in the amplified product using a solid phase carrier; performing single primer linear amplification using a third primer with the captured single strand as a template; performing exponential amplification using the third primer and the first primer, with the linearly amplified product as the template, to obtain a product containing the target regions.

    METHOD AND REAGENT FOR CONSTRUCTING NUCLEIC ACID DOUBLE-LINKER SINGLE-STRAND CYCLICAL LIBRARY
    6.
    发明申请
    METHOD AND REAGENT FOR CONSTRUCTING NUCLEIC ACID DOUBLE-LINKER SINGLE-STRAND CYCLICAL LIBRARY 审中-公开

    公开(公告)号：US20170355981A1

    公开(公告)日：2017-12-14

    申请号：US15529881

    申请日：2014-11-26

    申请人： BGI SHENZHEN ,  BGI SHENZHEN CO., LIMITED

    发明人： Yuan JIANG ,  Qiaoling LI ,  Andrei ALEXEEV ,  Evan HUROWITZ ,  Xia ZHAO ,  Tong WANG ,  Chao DONG ,  Dong LI ,  Radoje DRMANAC ,  Wenwei ZHANG ,  Hui JIANG

    IPC分类号： C12N15/10 ,  B01J19/00

    摘要： A method and reagent for constructing a nucleic acid double-linker single-strand cyclic library. The method comprises: breaking a nucleic acid into nucleic acid fragments; connecting a first linker sequence; producing by amplification a first product provided with the first linker sequence at either end, where a U nucleobase site is provided on primer sequences and a nicking enzyme recognition sequence is either provided or not provided on same, and a first affinity tag is provided on one of the primer sequences; using USER enzyme to cleave the first product; cyclizing the cleaved first product; treating the cyclization product with either a phosphatase or a nicking enzyme; using a solid-phase vector for combination with a cyclized molecule; performing a restrictive gap translation reaction; removing by digestion any portion that did not undergo the restrictive gap translation reaction; connecting a second linker sequence; producing by amplification a second product provided with the second linker sequence at either end; denaturing the second product, and cyclizing a single-strand nucleic acid molecule. The method allows an increase in the length of library insert fragments, a simplified library construction process, reduced library construction time, and reduced library construction costs.