METHOD FOR BREAKING NUCLEIC ACID AND ADDING ADAPTOR BY MEANS OF TRANSPOSASE, AND REAGENT

Title:

METHOD FOR BREAKING NUCLEIC ACID AND ADDING ADAPTOR BY MEANS OF TRANSPOSASE, AND REAGENT

Document Type and Number:

WIPO Patent Application WO/2016/058127

Kind Code:

A1

Abstract:

Provided are a method for breaking a nucleic acid and adding an adaptor by means of a transposase, and a reagent. The method comprises the following steps: conducting random breaking of a nucleic acid by using a transposase-embedded complex, wherein the transposase-embedded complex comprises a transposase and a first adaptor comprising a transposase identification sequence, and two ends of the broken nucleic acid are separately connected to the first adaptor and are separately provided with a gap; by means of purification or chemical reagent treatment, eliminating the influence of the transposase in the system on a follow-up reaction; connecting to a second adaptor at the gap by using a ligase, wherein a sequence of the second adaptor is different from a sequence of the first adaptor; and conducting a PCR reaction by using primers targeted to and combined with the first adaptor and the second adaptor respectively, so as to obtain a product whose two ends are respectively connected to different adaptor sequences.

Inventors:

GENG CHUNYU (CN)
CHEN RUOYING (CN)
GUO RONGRONG (CN)
ALEXEEV ANDREI (US)
ZHANG YINGXIN (US)
JIANG HUI (CN)
ZHANG WENWEI (CN)

Application Number:

PCT/CN2014/088542

Publication Date:

April 21, 2016

Filing Date:

October 14, 2014

Export Citation:

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Assignee:

BGI SHENZHEN CO LTD (CN)

International Classes:

C12N15/10; C12N15/11

Domestic Patent References:

WO2012103545A1	2012-08-02
WO2013131962A1	2013-09-12

Foreign References:

CN102264914A	2011-11-30
CN102943074A	2013-02-27
CN101125873A	2008-02-20
CN1145641A	1997-03-19
CN102703426A	2012-10-03
CN103710323A	2014-04-09

Other References: