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    Method and reagent for constructing nucleic acid double-linker single-strand cyclical library
    1.
    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

    Applicant: BGI SHENZHEN ,  BGI SHENZHEN CO., LIMITED

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

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

    Abstract: 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.

    Method for preparing nucleic acid library, its uses and kits
    3.
    发明授权
    Method for preparing nucleic acid library, its uses and kits 有权
    制备核酸文库的方法及其用途和试剂盒

    公开(公告)号：US09359642B2

    公开(公告)日：2016-06-07

    申请号：US14352492

    申请日：2012-10-16

    Applicant: BGI Shenzhen Co., Limited ,  BGI Shenzhen

    Inventor： Xuyang Yin ,  Chunlei Zhang ,  Hui Jiang ,  Xiuqing Zhang ,  Shengpei Chen

    IPC: C12Q1/68 ,  C12N15/10

    CPC classification number: C12Q1/6874 ,  C12N15/1093 ,  C12Q1/6853 ,  C12Q1/686 ,  C12Q1/6869 ,  C12Q2523/125 ,  C12Q2525/179 ,  C12Q2525/191 ,  C12Q2531/113 ,  C12Q2537/159

    Abstract: Provided are a method of constructing a nucleic acid library, a method of determining a nucleic acid sequence of a nucleic acid sample, and a kit thereof. The method of constructing the nucleic acid library includes the following steps: subjecting a nucleic acid sample to a DOP-PCR amplification, to obtain a first PCR amplification product; subjecting the first PCR amplification product to a second PCR amplification using a DOP-Amp primer, to obtain a second PCR amplification product; and subjecting the second PCR amplification product to an adaptor-ligation PCR, to obtain a third PCR amplification product, wherein the third PCR amplification product constitutes the nucleic acid library.

    Abstract translation: 提供构建核酸文库的方法，确定核酸样品的核酸序列的方法及其试剂盒。 构建核酸文库的方法包括以下步骤：使核酸样品进行DOP-PCR扩增，得到第一PCR扩增产物; 使用DOP-Amp引物对第一PCR扩增产物进行第二次PCR扩增，得到第二PCR扩增产物; 并对第二PCR扩增产物进行衔接连接PCR，得到第三PCR扩增产物，其中第三PCR扩增产物构成核酸文库。

    Method for preparing nucleic acid library, its uses and kits
    4.
    发明申请
    Method for preparing nucleic acid library, its uses and kits 有权
    制备核酸文库的方法及其用途和试剂盒

    公开(公告)号：US20140296084A1

    公开(公告)日：2014-10-02

    申请号：US14352492

    申请日：2012-10-16

    Applicant: BGI Shenzhen Co., Limited ,  BGI Shenzhen

    Inventor： Xuyang Yin ,  Chunlei Zhang ,  Hui Jiang ,  Xiuqing Zhang ,  Shengpei Chen

    IPC: C12Q1/68

    CPC classification number: C12Q1/6874 ,  C12N15/1093 ,  C12Q1/6853 ,  C12Q1/686 ,  C12Q1/6869 ,  C12Q2523/125 ,  C12Q2525/179 ,  C12Q2525/191 ,  C12Q2531/113 ,  C12Q2537/159

    Abstract: Provided are a method of constructing a nucleic acid library, a method of determining a nucleic acid sequence of a nucleic acid sample, and a kit thereof. The method of constructing the nucleic acid library includes the following steps: subjecting a nucleic acid sample to a DOP-PCR amplification, to obtain a first PCR amplification product; subjecting the first PCR amplification product to a second PCR amplification using a DOP-Amp primer, to obtain a second PCR amplification product; and subjecting the second PCR amplification product to an adaptor-ligation PCR, to obtain a third PCR amplification product, wherein the third PCR amplification product constitutes the nucleic acid library.

    Abstract translation: 提供构建核酸文库的方法，确定核酸样品的核酸序列的方法及其试剂盒。 构建核酸文库的方法包括以下步骤：使核酸样品进行DOP-PCR扩增，得到第一PCR扩增产物; 使用DOP-Amp引物对第一PCR扩增产物进行第二次PCR扩增，得到第二PCR扩增产物; 并对第二PCR扩增产物进行衔接连接PCR，得到第三PCR扩增产物，其中第三PCR扩增产物构成核酸文库。

    Method of constructing sequencing library
    5.
    发明授权
    Method of constructing sequencing library 有权

    公开(公告)号：US10456769B2

    公开(公告)日：2019-10-29

    申请号：US15667841

    申请日：2017-08-03

    Applicant: BGI Shenzhen ,  BGI Shenzhen Co., Limited

    Inventor： Ou Wang ,  Xiaofang Cheng ,  Liangying Zou ,  Cankun Chang ,  Hui Jiang ,  Wenwei Zhang

    IPC: B01J19/00 ,  C12N15/10

    Abstract: Provided is a method of constructing a sequencing library. The method includes 1) providing a single-stranded DNA fragment from a biological sample; 2) subjecting the single-stranded DNA fragment to whole genomic amplification to obtain a whole genome amplification product; 3) fragmenting the whole genome amplification product using a transposase embedded with two adaptors to obtain a fragmented product with two adaptors respectively at two ends; and 4) amplifying the fragmented product with two adaptors respectively at two ends using a tag sequence and a pair of primers to obtain said sequencing library.

    Target region enrichment method based on multiplex PCR, and reagent
    6.
    发明授权
    Target region enrichment method based on multiplex PCR, and reagent 有权

    公开(公告)号：US10435736B2

    公开(公告)日：2019-10-08

    申请号：US15537396

    申请日：2015-12-09

    Applicant: BGI SHENZHEN ,  BGI SHENZHEN CO., LIMITED

    Inventor： Jing Guo ,  Rongrong Guo ,  Meiyan Li ,  Chunyu Geng ,  Hui Jiang

    IPC: C12Q1/68 ,  C12P19/34 ,  C12Q1/6806 ,  C12Q1/6855

    Abstract: 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 a sequence combination for producing nucleic acid fragments
    8.
    发明授权
    Method and a sequence combination for producing nucleic acid fragments 有权

    公开(公告)号：US10344317B2

    公开(公告)日：2019-07-09

    申请号：US15518760

    申请日：2014-10-13

    Applicant: BGI SHENZHEN CO., LIMITED

    Inventor： Hongyan Han ,  Chunyu Geng ,  Guanying Guo ,  Wenwei Zhang ,  Hui Jiang ,  Yuan Jiang

    IPC: C12P19/34 ,  C12Q1/6806 ,  C12Q1/6855 ,  C12Q1/686

    Abstract: Disclosed are a nucleic acid fragmentation method and a sequence combination. The method comprises the following steps: subjecting a denatured nucleic acid to annealing and an extension reaction by using a single-stranded 5′-end extension primer, wherein the single-stranded 5′-end extension primer comprises a sequencing platform adaptor sequence of a 5′ end and a connected random sequence, and the random sequence is subjected to annealing on a random site of the denatured nucleic acid; and directionally connecting a double-stranded 3′-end adaptor sequence to the 3′ end of the nucleic acid generated in the extension reaction, and carrying out denaturalization and purification to obtain a fragmented single-stranded nucleic acid with adaptor sequences on two ends.

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