PCR primer and application thereof in DNA fragment connection

A primer sequence and product technology, applied in DNA/RNA fragments, DNA preparation, recombinant DNA technology, etc., can solve the problems of short sequence splicing that cannot provide long transcripts, loss of alternative splicing, and short sequencing sequences

Active Publication Date: 2020-10-09
BGI TECH SOLUTIONS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sequencing sequence of the second-generation sequencing technology is short, and short-sequence splicing cannot provide a large number of long transcripts and loses important information such as alternative splicing. Therefore, the de novo sequencing of the transcriptome starts with the third-generation sequencing technology of PacBio

Method used

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  • PCR primer and application thereof in DNA fragment connection
  • PCR primer and application thereof in DNA fragment connection
  • PCR primer and application thereof in DNA fragment connection

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Embodiment Construction

[0030] Embodiments of the invention are described in detail below, examples of which are illustrated in the accompanying drawings. The embodiments described below by referring to the figures are exemplary and are intended to explain the present invention and should not be construed as limiting the present invention.

[0031] Aiming at the problem that the existing full-length transcript library building method cannot make full use of the advantages of the enzyme read length of the Pacbio platform, the present invention designs a full-length transcriptome end-to-end library construction scheme, and connects the PCR products of the transcriptome end-to-end through sticky ends. Thereby increasing the average length of the library, so that more transcript data can be obtained after sequencing.

[0032]According to the embodiment of the present invention, the reverse transcription step is consistent with the conventional library construction, and the template switching principle is...

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Abstract

The invention provides a PCR primer and an application thereof in DNA fragment connection. The PCR primer comprises a palindrome sequence segment and any PCR primer sequence segment, the 3'end of thepalindrome sequence segment is connected with the 5'end of the any PCR primer sequence segment, the nucleotide of the 3' end of the palindrome sequence segment is U, and the any PCR primer sequence segment is complementarily paired with the 3'end of a pre-amplified DNA template. The primer provided by the embodiment of the invention can be used for constructing a sequencing library; in the construction process, the primer provided by the embodiment of the invention is used as a PCR amplification primer based on DNA, the two ends of an obtained amplification product DNA double strand have a palindrome sequence structure, and the 3'end of the palindrome sequence structure has nucleotide U, so that a foundation is laid for subsequent interconnection of different amplification product double strand DNAs.

Description

technical field [0001] The present invention relates to the field of biotechnology. Specifically, the present invention relates to PCR primers and their application in DNA fragment ligation. More specifically, the present invention relates to PCR primers, PCR amplification methods, methods for constructing sequencing libraries, and full-length transcriptomes. sequencing method. Background technique [0002] Genome and transcriptome sequencing is fundamental work in the life sciences. Since the vast majority of non-model organisms lack genomic data, full-length transcriptome sequencing has become particularly important. Full-length transcripts can greatly promote basic and applied research on gene function, gene expression regulation, and evolutionary relationships in these species. Currently, the vast majority of transcriptome data are obtained based on second-generation high-throughput sequencing technologies. However, the sequencing sequence of the second-generation sequ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12Q1/6869C12N15/10C12N15/11
CPCC12N15/1093C12Q1/686C12Q1/6869
Inventor 陈智超唐冲阮凤英郭梅石卓兴杨林峰
Owner BGI TECH SOLUTIONS
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