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Construction method and sequencing method of genetic typing sequencing library

A technology for genotyping, sequencing, and construction methods, which is applied in chemical libraries, biochemical equipment and methods, and microbial determination/inspection, etc. problem, to achieve the effect of improving the effective amount, improving the efficiency of building the library, and increasing the length

Active Publication Date: 2015-04-29
BEIJING NOVOGENE TECH CO LTD
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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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  • Construction method and sequencing method of genetic typing sequencing library
  • Construction method and sequencing method of genetic typing sequencing library
  • Construction method and sequencing method of genetic typing sequencing library

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[0021] It should be noted that, in the case of no conflict, the embodiments in the present application and the features in the embodiments can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and examples.

[0022] In view of the cumbersome process of building a library in the construction method of the genotyping sequencing library in the prior art, the time is long, and the quality of the sequencing data obtained by sequencing the constructed library is not high, resulting in data waste and other shortcomings, in a typical method of the present invention In an embodiment, a method for constructing a genotyping sequencing library is provided, such as figure 1 As shown, the method includes the following steps: S1, when the genomic DNA is digested for the first time to generate the first restriction fragment, at the same time, a sample tag sequence is connected to both ends of the first restriction ...

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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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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/10C40B50/06
CPCC12Q1/6869C40B50/06C12Q2535/122
Inventor 王大伟刘运超蒋智李明渊朱海浩孙晴晴
Owner BEIJING NOVOGENE TECH CO LTD
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