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A primer set and method for constructing an amplicon library suitable for single-end sequencing

An amplicon library and primer set technology, which is applied in the field of amplicon library construction primer set and construction, can solve the problem that the amplicon library cannot meet the requirements of single-end sequencing, and can shorten the TAT time, reduce the sequencing time, and facilitate the The effect of promoting the application

Active Publication Date: 2022-04-01
臻和(北京)生物科技有限公司 +1
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Problems solved by technology

[0007] Based on the problem that the amplicon library in the prior art cannot meet the requirements of single-end sequencing, the present invention has developed a primer set and library construction method for the amplicon library that can meet the requirements of single-end sequencing

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  • A primer set and method for constructing an amplicon library suitable for single-end sequencing
  • A primer set and method for constructing an amplicon library suitable for single-end sequencing
  • A primer set and method for constructing an amplicon library suitable for single-end sequencing

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Embodiment 1

[0101] Taking 4 FFPEDNA samples (the sample numbers are DS0202, DS0203, DS0236 and DS0251 respectively, all of which are non-small cell lung cancer samples) as the detection objects, the amplicon libraries of the 4 samples were constructed using the primer set provided by the present invention, and a single End-to-end sequencing, the specific process is as follows:

[0102] 1. Primer panel design

[0103] (1) Design of multiple primers for the first round of PCR

[0104] For each target amplicon, design 2 pairs of multiplex primers, each pair of multiplex primers contains 1 multiplexed upstream primer and 1 multiplexed downstream primer;

[0105] Wherein, the first multiple upstream primers are the first adapter sequence and the target amplicon-specific primer sequence according to the 5'-3' direction, and the first multiple downstream primers are the second adapter sequence according to the 5'-3' direction and target amplicon downstream specific primer sequences;

[0106] ...

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Abstract

The invention relates to the technical field of high-throughput sequencing, in particular to an amplicon library construction primer set and a construction method suitable for single-end sequencing. The primer set provided by the present invention includes at least one multiplex primer pair of target amplicon; for each target amplicon, the multiplex primer pair includes 2 pairs of multiplex primers, and each pair of multiplex primers includes one multiplex upstream primer and one A multiplex downstream primer; the multiplex upstream primer and the multiplex downstream primer both contain a linker sequence and a target amplicon-specific primer sequence; in the two pairs of multiplex primers, the two multiplex upstream primers contain different linker sequences, and two multiplex upstream primers contain different linker sequences. The linker sequences contained in the downstream primers are also different. The primer set can be used to construct an amplicon library suitable for single-end sequencing, which effectively reduces the sequencing time and TAT time.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing, in particular to an amplicon library construction primer set and construction method suitable for single-end sequencing. Background technique [0002] Amplicon library construction is a library construction method that adds adapters to both ends of the DNA fragments to be tested through PCR reactions. Only two rounds of PCR and two steps of purification are required to obtain the library of the target region. The first step is to contain the target region-specific The primers of the sex-binding sequence bind to the target region, and the second step is to connect the sequencing adapter through the PCR reaction, so that the constructed library can be used for on-machine sequencing. Compared with the capture library construction method, the amplicon library construction completes the library construction by customizing the primer panel. In clinical applications, the method of am...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C40B50/06C12N15/11C12Q1/6869
CPCC40B50/06C12Q1/6869C12Q2531/113C12Q2537/143C12Q2535/122
Inventor 李凤明魏丽许青陈维之杜波
Owner 臻和(北京)生物科技有限公司
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