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.

公开(公告)号：US20180044667A1

公开(公告)日：2018-02-15

申请号：US15510904

申请日：2014-10-14

Applicant: BGI SHENZHEN CO., LIMITED

Inventor： Chunyu Geng ,  Rongrong Guo ,  Ruoying Chen ,  Lingyu He ,  Wenwei Zhang ,  Hui Jiang

IPC: C12N15/10 ,  C12N11/06 ,  C12N9/00 ,  C40B50/14 ,  C12N15/66 ,  C12Q1/68 ,  C40B50/06 ,  C40B40/08 ,  C12N9/22

CPC classification number: C12Q1/6853 ,  B01J19/0046 ,  B01J2219/00529 ,  C12N9/22 ,  C12N9/93 ,  C12N11/06 ,  C12N15/10 ,  C12N15/1065 ,  C12N15/1082 ,  C12N15/1093 ,  C12N15/66 ,  C12Q1/6806 ,  C12Q1/6811 ,  C12Q1/6837 ,  C12Q1/6855 ,  C12Q1/686 ,  C12Q1/6874 ,  C12Q2525/191 ,  C12Q2565/537 ,  C40B40/06 ,  C40B40/08 ,  C40B50/06 ,  C40B50/14 ,  C40B60/14 ,  C12Q2521/525 ,  C12Q2535/122 ,  C12N15/1089 ,  C12Q2525/186 ,  C12Q2521/101

Abstract: Provided in the present invention are a method for constructing a nucleic acid single-stranded cyclic library and reagents thereof. The method comprises the steps of using a transposase embedding complex to randomly break nucleic acids and connect a first linker; connecting a second linker at a gap; performing a first PCR reaction, wherein the 5′ end of one of the primers has a first affinity tag, resulting in a product with two ends connected to different linker sequences; binding the product to a solid vector having a second affinity tag; degenerating and separating single strands having no affinity tag; and cyclizing the single strands.

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：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：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：US10023906B2

公开(公告)日：2018-07-17

申请号：US15510904

申请日：2014-10-14

Applicant: BGI SHENZHEN CO., LIMITED

Inventor： Chunyu Geng ,  Rongrong Guo ,  Ruoying Chen ,  Lingyu He ,  Wenwei Zhang ,  Hui Jiang

IPC: C12Q1/68 ,  C12N9/22 ,  C12Q1/6853 ,  C12N15/10 ,  C40B40/08 ,  C12N11/06 ,  C12N9/00 ,  C12Q1/686 ,  C40B50/06 ,  C40B50/14 ,  C12Q1/6806 ,  C12N15/66

CPC classification number: C12Q1/6853 ,  B01J19/0046 ,  B01J2219/00529 ,  C12N9/22 ,  C12N9/93 ,  C12N11/06 ,  C12N15/10 ,  C12N15/1065 ,  C12N15/1082 ,  C12N15/1093 ,  C12N15/66 ,  C12Q1/6806 ,  C12Q1/6811 ,  C12Q1/6837 ,  C12Q1/6855 ,  C12Q1/686 ,  C12Q1/6874 ,  C12Q2525/191 ,  C12Q2565/537 ,  C40B40/08 ,  C40B50/06 ,  C40B50/14 ,  C40B60/14 ,  C12Q2521/525 ,  C12Q2535/122 ,  C12N15/1089 ,  C12Q2525/186 ,  C12Q2521/101

Abstract: Provided in the present invention are a method for constructing a nucleic acid single-stranded cyclic library and reagent kit thereof. The method comprises the steps of using a transposase embedding complex to randomly break nucleic acids and connect a first linker; connecting a second linker at a gap; performing a first PCR reaction, wherein the 5′ end of one of the primers has a first affinity tag, resulting in a product with two ends connected to different linker sequences; binding the product to a solid vector having a second affinity tag; degenerating and separating single strands having no affinity tag; and cyclizing the single strands.

公开(公告)号：US09890375B2

公开(公告)日：2018-02-13

申请号：US15510877

申请日：2014-09-12

Applicant: BGI SHENZHEN CO., LIMITED

Inventor： Chunyu Geng ,  Dennis G. Ballinger ,  Yanyan Zhang ,  Shujin Fu ,  Lingyu He ,  Wenwei Zhang ,  Hui Jiang

IPC: C12Q1/68 ,  C12N15/10 ,  B01J19/00 ,  C40B60/14

CPC classification number: C12Q1/6853 ,  B01J19/0046 ,  B01J2219/00529 ,  C12N9/22 ,  C12N9/93 ,  C12N11/06 ,  C12N15/10 ,  C12N15/1065 ,  C12N15/1082 ,  C12N15/1093 ,  C12N15/66 ,  C12Q1/6806 ,  C12Q1/6811 ,  C12Q1/6837 ,  C12Q1/6855 ,  C12Q1/686 ,  C12Q1/6874 ,  C12Q2525/191 ,  C12Q2565/537 ,  C40B40/06 ,  C40B40/08 ,  C40B50/06 ,  C40B50/14 ,  C40B60/14 ,  C12Q2521/525 ,  C12Q2535/122 ,  C12N15/1089 ,  C12Q2525/186 ,  C12Q2521/101

Abstract: Provided are an isolated oligonucleotide and a use thereof in nucleic acid sequencing, wherein the isolated oligonucleotide comprises a first strand, wherein the 5′-end nucleotide of the first strand has a phosphate group, and the 3′-end nucleotide of the first strand is a dideoxynucleotide, and a second strand, wherein the 5′-end nucleotide of the second strand does not have a phosphate group, and the 3′-end nucleotide of the second strand is a dideoxynucleotide, wherein the first strand is longer than the second strand in length, and a double-stranded structure is formed between the first strand and the second strand.

公开(公告)号：US20170275609A1

公开(公告)日：2017-09-28

申请号：US15510877

申请日：2014-09-12

Applicant: BGI SHENZHEN CO., LIMITED

Inventor： Chunyu Geng ,  Dennis G. Ballinger ,  Yanyan Zhang ,  Shujin Fu ,  Lingyu He ,  Wenwei Zhang ,  Hui Jiang

IPC: C12N15/10 ,  C40B60/14 ,  C12Q1/68 ,  B01J19/00

CPC classification number: C12Q1/6853 ,  B01J19/0046 ,  B01J2219/00529 ,  C12N9/22 ,  C12N9/93 ,  C12N11/06 ,  C12N15/10 ,  C12N15/1065 ,  C12N15/1082 ,  C12N15/1093 ,  C12N15/66 ,  C12Q1/6806 ,  C12Q1/6811 ,  C12Q1/6837 ,  C12Q1/6855 ,  C12Q1/686 ,  C12Q1/6874 ,  C12Q2525/191 ,  C12Q2565/537 ,  C40B40/06 ,  C40B40/08 ,  C40B50/06 ,  C40B50/14 ,  C40B60/14 ,  C12Q2521/525 ,  C12Q2535/122 ,  C12N15/1089 ,  C12Q2525/186 ,  C12Q2521/101

Abstract: Provided are an isolated oligonucleotide and a use thereof in nucleic acid sequencing, wherein the isolated oligonucleotide comprises a first strand, wherein the 5′-end nucleotide of the first strand has a phosphate group, and the 3′-end nucleotide of the first strand is a dideoxynucleotide, and a second strand, wherein the 5′-end nucleotide of the second strand does not have a phosphate group, and the 3′-end nucleotide of the second strand is a dideoxynucleotide, wherein the first strand is longer than the second strand in length, and a double-stranded structure is formed between the first strand and the second strand.

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.