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.

公开(公告)号：US20170356039A1

公开(公告)日：2017-12-14

申请号：US15510890

申请日：2014-11-21

Applicant: BGI SHENZHEN CO., LIMITED

Inventor： Yuan Jiang ,  Jing Guo ,  Xiaojun Ji ,  Chunyu Geng ,  Kai Tian ,  Xia Zhao ,  Huaiqian Xu ,  Wenwei Zhang ,  Hui Jiang ,  Radoje Drmanac

IPC: C12Q1/68 ,  C12N15/10

Abstract: Provided are a vesicular linker and a single-chain cyclic library constructed by using the linker. The library can be used for RNA sequencing and other sequencing platforms dependent on a single-stranded cyclic library, and has the advantages of high throughput sequencing, high accuracy and simple operations.

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.

公开(公告)号：US10563196B2

公开(公告)日：2020-02-18

申请号：US15519530

申请日：2014-10-17

Applicant: BGI SHENZHEN

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

IPC: C12N15/10 ,  C12Q1/6806 ,  C12Q1/68 ,  C12Q1/6853 ,  C12Q1/686 ,  C40B40/08 ,  C40B50/06

Abstract: The present invention provides a primer for nucleic acid random fragmentation and a nucleic acid random fragmentation method. The primer consists of a plurality of upstream random primers and downstream random primers. The sequence composition of the upstream random primers is 5′-X-Y-3′, and the sequence composition of the downstream random primers is 5′-P-Y′-X′-close-3′, wherein Y and Y′ are random sequences, X is all or part of sequences of a sequencing platform 5′ end adaptor, X′ is all or part of sequences of a sequencing platform 3′ end adaptor, P is phosphorylation modification, and close is close modification. The primer of the present invention adopts double random anchoring of both the upstream random primers and the downstream random primers, and a DNA sample can be randomly broken.

公开(公告)号：US20180030532A1

公开(公告)日：2018-02-01

申请号：US15528362

申请日：2014-11-21

Applicant: BGI Shenzhen Co., Limited

Inventor： Yuan Jiang ,  Xia Zhao ,  Qiaoling Li ,  Shengmao Liu ,  Bo Wang ,  Li Chen ,  Wenwei Zhang ,  Hui Jiang ,  Radoje Drmanac

IPC: C12Q1/68

CPC classification number: C12Q1/6874 ,  C12Q1/68 ,  C12Q1/6855 ,  C12Q1/6869 ,  C12Q2525/191 ,  C12Q2525/301 ,  C12Q2535/122

Abstract: Provided are an adaptor element in a bubble shape, a method of constructing a sequencing library with such an adapter element. The adaptor element is a hybrid formed with a long-chain nucleic acid A and a short-chain nucleic acid B. The hybrid is in the bubble shape with paired regions at two terminals and a non-paired region in the middle.

公开(公告)号：US20180291371A1

公开(公告)日：2018-10-11

申请号：US15510882

申请日：2014-11-26

Applicant: BGI SHENZHEN CO., LIMITED

Inventor： Chunyu Geng ,  Ruoying Chen ,  Yuan Jiang ,  Xia Zhao ,  Rongrong Guo ,  Lingyu He ,  Yaqiao Li ,  Wenwei Zhang ,  Hui Jiang ,  Radoje Drmanac

IPC: C12N15/10 ,  C12Q1/6806

CPC classification number: C12N15/1093 ,  C12N15/1089 ,  C12N15/66 ,  C12Q1/6806 ,  C12Q1/6853 ,  C12Q1/6855 ,  C12Q2525/191 ,  C12Q2521/307 ,  C12Q2521/507 ,  C12Q2525/121 ,  C12Q2525/307 ,  C12Q2563/131

Abstract: Provided are a method for constructing a nucleic acid single-stranded cyclic library and the reagents used therein. By the combination of interruption via a transposase with a restricted nick translation reaction, the method realizes a simple and rapid nucleic acid single-stranded cyclic library construction.