Invention Grant
- Patent Title: Method and reagent for constructing nucleic acid double-linker single-strand cyclical library
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Application No.: US15529881Application Date: 2014-11-26
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Publication No.: US10479991B2Publication Date: 2019-11-19
- Inventor: Yuan Jiang , Qiaoling Li , Andrei Alexeev , Evan Hurowitz , Xia Zhao , Tong Wang , Chao Dong , Dong Li , Radoje Drmanac , Wenwei Zhang , Hui Jiang
- Applicant: BGI SHENZHEN , BGI SHENZHEN CO., LIMITED
- Applicant Address: CN Shenzhen, Guangdong
- Assignee: MGI TECH CO., LTD
- Current Assignee: MGI TECH CO., LTD
- Current Assignee Address: CN Shenzhen, Guangdong
- Agency: Ladas & Parry, LLP
- International Application: PCT/CN2014/092296 WO 20141126
- International Announcement: WO2016/082129 WO 20160602
- Main IPC: C12N15/10
- 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.
Public/Granted literature
- US20170355981A1 METHOD AND REAGENT FOR CONSTRUCTING NUCLEIC ACID DOUBLE-LINKER SINGLE-STRAND CYCLICAL LIBRARY Public/Granted day:2017-12-14
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