Method and kit for constructing sequencing library

Abstract

The invention provides a method and a kit for constructing a sequencing library. The method comprises the following steps: performing enzyme digestion on a genome DNA of a sample to be tested to obtain an enzyme digestion fragment with sticky terminals; adding P1 connectors at the two ends of the enzyme digestion fragment to obtain a fragment with the P1 connectors; fragmenting the fragment with the P1 connectors to obtain fragments with the target sizes; adding P2 connectors at the two ends of the fragments with the target sizes to obtain a sequencing library, wherein the genome DNA comprises a recognition sequence which can be recognized by an enzyme; the recognition sequence comprises a palindromic sequence consisting of basic groups at the two ends of the recognition sequence, and a variable sequence in the middle of the palindromic sequence; the variable sequence comprises a one or more basic groups; the sticky terminals comprise one or more basic groups in the variable sequence. Through enzyme digestion on the enzyme of the variable sequence in the recognition sequence, different capturing tag numbers can be achieved through the types and the number of the basic groups in the fixing connectors, so that the flexibility is improved.

Description

technical field [0001] The invention relates to the field of high-throughput sequencing, in particular to a method for constructing a sequencing library and a kit for constructing the sequencing library. Background technique [0002] Restriction-site Associated DNA (RAD)-based sequencing technology, that is, RAD-seq technology is a simplified genome sequencing based on whole-genome restriction sites developed on the basis of next-generation sequencing. technology. The method has a simple technical process and is not limited by the presence or absence of a reference genome. It can greatly simplify the complexity of the genome and reduce the cost of experiments. Tens of thousands of polymorphic markers can be obtained through a single sequencing. At present, RAD-seq technology has been successfully applied to the construction of ultra-high-density genetic maps, fine mapping of important traits, auxiliary genome sequence assembly, population genomics and phylogenetics and othe...

AI Technical Summary

Problems solved by technology

[0008] The main purpose of the present invention is to provide a method for constructing a sequencing library and a kit for constructing a sequencing library, so as to solve the problems in the prior art when constructing a sequencing (RAD-Seq) library based on restriction enzyme site-associated DNA. There are problems of poor flexibility and cost waste

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