Construction method and sequencing method of genetic typing sequencing library

Abstract

The invention discloses a construction method and a sequencing method of a genetic typing sequencing library. The construction method comprises the following steps: S1, bonding sample label sequences at the two ends of a first enzyme digestion fragment while a gene group DNA is subjected to enzyme digestion for the first time to produce the first enzyme digestion fragment so as to obtain a first enzyme digestion fragment with a label; S2, performing PCR amplification on the first enzyme digestion fragment by utilizing an amplification primer with a library label sequence to obtain the gene typing sequencing library. According to the construction method, the sample label sequences are directly bonded at the two ends of the first enzyme digestion fragment while the gene group DNA is subjected to enzyme digestion for the first time to produce the first enzyme digestion fragment, so that the constructed library can directly get into the next step of reading a target enzyme digestion fragment sequence after the sample label information is read during sequencing by a computer, and the reading length of the target enzyme digestion fragment sequence is increased, the invalid data volume is reduced, and the effective quantity of sequencing data is increased.

Description

technical field [0001] The present invention relates to the field of high-throughput sequencing, in particular to a method for constructing a genotyping sequencing library and a sequencing method. Background technique [0002] GBS (Genotyping-by-Sequencing) technology is mainly used in the development of molecular markers for samples, the construction of ultra-high-density genetic maps, population genetic analysis and population GWAS analysis. Based on SLAF-seq (Specific Length Amplified Fragments sequencing) is a large-scale sample genotyping method, which is a simplified genome sequencing method based on genome-wide restriction sites developed on the basis of next-generation sequencing. The basic principle of SLAF-seq is: use restriction endonucleases to digest genomic DNA samples, reduce genome complexity and perform high-throughput sequencing, and then perform marker development, genetic map drawing, and genome-wide association analysis on large-scale samples Wait. [...

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Problems solved by technology

[0004] However, the construction method of the above-mentioned SLAF-seq genotyping sequencing library in the prior art is not only cumbersome and takes a long time, but also the number of samples that can be mixed in the constructed library is limited, and the quality of the sequencing data obtained after sequencing is not good. High, often resulting in waste of sequencing data

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