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Method utilizing liquid phase capture to carry out gene typing of large yellow croaker genome

A technology of genotyping and large yellow croaker, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve the problems of high cost, difficulty in application, and complicated gene chip design, so as to improve efficiency, specificity, Effects of High Efficiency Genome Typing Technology

Inactive Publication Date: 2016-12-07
JIMEI UNIV
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AI Technical Summary

Problems solved by technology

However, the design of the gene chip is complex and expensive, and it is difficult to apply it to the field of high-throughput SNP / InDel genotyping at specific sites in large yellow croaker. There is an urgent need for a technology that can obtain specific genome fragments of large yellow croaker for reliable genotyping at low cost.

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  • Method utilizing liquid phase capture to carry out gene typing of large yellow croaker genome
  • Method utilizing liquid phase capture to carry out gene typing of large yellow croaker genome

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Embodiment 1

[0023] In the embodiment of the present invention, a liquid-phase capture method for whole-genome genotyping of large yellow croaker:

[0024] 1. Resequencing data using large yellow croaker

[0025] In view of the fact that the above-mentioned individuals used for genome sequencing are highly homozygous individuals with few heterozygous sites / SNP sites, 100 large yellow croakers (50 male and 50 male) were selected from different breeding groups in September 2012 , Genome resequencing was carried out by constructing a 300bp small fragment library. The two groups of resequencing obtained a total of 20Gb of raw data, and the average sequencing depth reached 26.8.

[0026] 2. Determine the capture site, probe design

[0027] The sequencing results were compared to the above draft genome for SNP marker mining. A total of 12,161,573 SNP sites with a sequencing depth greater than 5X, 505,286 on exons, 4,949,058 on introns, and 5,135,488 between genes were obtained. According to ...

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Abstract

The invention discloses a method utilizing liquid phase capture to carry out gene typing of large yellow croaker genome. The method comprises the following steps: subjecting a large yellow croaker to whole genome re-sequencing, at the same time, carrying out large yellow croaker whole genome SNP / InDel mining to provide high quality SNP labeled sites; screening and utilizing SNP labels obtained by large yellow croaker whole genome re-sequencing to design probes; utilizing the large yellow croaker whole genome to establish a large yellow croaker genome library; utilizing a liquid phase to capture the large yellow croaker genome sections; and subjecting the captured large yellow croaker genome sections to high throughput sequencing and SNP / InDel gene typing. The provided method effectively solves the problem that a whole gene simplified genome typing technology may cause unevenness of label density and most labels are originated from non-coding regions. According to the method, four probes are designed to effectively improve the specificity of hybrid of sequences where each SNP stays, the efficiency of section capture is improved; and the method has an important application in the large scale typing of large yellow croakers with special SNP labels.

Description

technical field [0001] The invention relates to the technical field of genotyping, in particular to a method for genotyping the large yellow croaker genome by liquid-phase capture. Background technique [0002] Molecular markers refer to the characteristic differences in genome sequences of organisms or populations. Single nucleotide polymorphism (SNP) and short insertion and deletion (InDel) are polymorphic sites among individuals caused by one or several base differences on the biological genome, and are an important class of molecular markers. SNP is considered by many people to be the best marker selection in the future. It has the characteristics of stable inheritance, wide distribution, easy estimation of allele frequency, important biological significance, and suitable for large-scale detection. At present, SNP / InDel markers have become an important tool for identification of biological populations, analysis of genetic structure, construction of linkage maps, linkage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 肖世俊刘洋王志勇
Owner JIMEI UNIV
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