Title:
METHOD FOR PREPARING NESTED MULTIPLEX PCR HIGH-THROUGHPUT SEQUENCING LIBRARY AND KIT
Document Type and Number:
WIPO Patent Application WO/2020/258084
Kind Code:
A1
Abstract:
Provided are a method for preparing a nested multiplex PCR high-throughput sequencing library and a kit, wherein the method for preparing the nested multiplex PCR high-throughput sequencing library comprises: amplifying a target region by using a forward primer and a reverse primer, wherein the forward primer is a forward specific sequence, and the reverse primer comprises a 3' end reverse specific sequence and a 5' end first universal sequencing sequence; purifying a product of a first round of PCR amplification; amplifying the purified product by using a nested primer, a first label primer and a second universal primer, wherein the nested primer is located downstream of the forward primer, the nested primer comprises a 5' end second universal sequencing sequence and a 3' end specific sequence, the 3' end sequence of the first label primer is the same as part or all of the 5' end sequence of the reverse primer, and the first label primer further comprises a first label sequence, the 3' end sequence of a second universal primer is the same as part or all of 5' end sequence of the nested primer. The method for preparing the nested multiplex PCR high-throughput sequencing library can effectively avoid dimers generated in the multiplex PCR amplification process, and the nested amplification method can greatly increase the specificity of PCR amplification.
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Inventors:
YANG LIN (CN)
ZHANG YANYAN (CN)
CHEN FANG (CN)
JIANG HUI (CN)
ZHANG YANYAN (CN)
CHEN FANG (CN)
JIANG HUI (CN)
Application Number:
PCT/CN2019/093066
Publication Date:
December 30, 2020
Filing Date:
June 26, 2019
Export Citation:
Assignee:
MGI TECH CO LTD (CN)
International Classes:
C12Q1/6869; C12Q1/6844
Domestic Patent References:
| WO2018112806A1 | 2018-06-28 | |||
| WO2018232597A1 | 2018-12-27 |
Foreign References:
| CN101988124A | 2011-03-23 | |||
| CN107312847A | 2017-11-03 | |||
| CN1990879A | 2007-07-04 | |||
| US20130045894A1 | 2013-02-21 |
Other References:
See also references of EP 3992303A4
Attorney, Agent or Firm:
DHC IP ATTORNEYS (CN)
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