Double-stranded linker, its application and method for constructing paired-end dna library

Abstract

The invention discloses a double chain adaptor, application thereof and a method for constructing end-pairing DNA library. The double chain adaptor comprises an adaptor A and an adaptor B, and the adaptor A and the adaptor B respectively comprise a LoxP sequence and an auxiliary sequence for linking the LoxP sequence to the two ends of a to-be-cyclized DNA fragment in a same direaction. By applying the technical scheme provided by the invention, based on the characteristics of the Cre recombinase / LoxP action system (when two LoxP sequences are disposed at one DNA chain and has the same direction, Cre recombinase is capable of effectively excising the sequence between the two LoxP sites, and the excised sequence automatically becomes annularity, so that the cyclization of a DNA fragment is realized), the double chain adaptor is designed and is efficiently linked to the two ends of a DNA fragment for constructing the end-pairing DNA library in a T-A clone manner, and the Cre recombinase is employed for efficiently enabling the DNA fragment for constructing the end-pairing DNA library to be cyclized.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing, in particular to a double-stranded linker, its application and a method for constructing a paired-end DNA library. Background technique [0002] Whole-genome Denovo sequencing is also called genome de novo sequencing. It does not rely on the existing sequence information, directly sequences the genome of a certain species, and uses bioinformatics methods to splice and assemble the sequences to obtain the genome of the species. Atlas. Through the systematic study of the whole genome sequence of a species, the genome and important functional gene sequence information of the species can be obtained, the evolution history of the species can be analyzed, and the molecular mechanism of growth and development, the generation of important traits and the adaptation to the environment of the species can be clarified. [0003] Before the advent of next-generation sequencing, the genome-w...

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Problems solved by technology

[0004] The present invention aims to provide a double-stranded linker, its application, and a method for constructing a paired-end DNA library, so as to solve the problem that the construction method of the Mate-paired DNA library in the prior art not only has low circularization efficiency, but also is difficult to effectively distinguish which sequences after sequencing Technical issues stemming from both ends of the junction

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