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SNP (single nucleotide polymorphism) marker for distinguishing Chinese population and Japan population of haliotis discus hannai

A wrinkled abalone and colony technology, applied in recombinant DNA technology, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effects of simple operation, low cost and high accuracy

Active Publication Date: 2015-12-30
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of hybrid breeding, it is difficult to distinguish the Japanese population purchased at a high price from the Chinese population morphologically. Therefore, the development of molecular markers that can identify the two populations is very important to determine the parental source of hybrid breeding, but there is no relevant research. to report

Method used

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  • SNP (single nucleotide polymorphism) marker for distinguishing Chinese population and Japan population of haliotis discus hannai
  • SNP (single nucleotide polymorphism) marker for distinguishing Chinese population and Japan population of haliotis discus hannai
  • SNP (single nucleotide polymorphism) marker for distinguishing Chinese population and Japan population of haliotis discus hannai

Examples

Experimental program
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Embodiment 1

[0025] Using next-generation sequencing technology, high-throughput sequencing was performed on 5 Chinese individuals and 5 Japanese individuals of Abalone rugosa, and the sequencing reads of all individuals were used to construct the genome scaffold of Abalone rugosa, which was used as a reference genome sequence for subsequent analysis. Compare the sequencing reads of Chinese individuals and Japanese individuals to the reference genome, screen the SNP sites and calculate their allele frequencies in the Chinese and Japanese populations, and use the Chi-squared test to calculate the alleles of the two populations The difference in frequency is used to screen the SNP sites with significant differences. A total of 5 SNP sites were screened out, wherein the site HdhSNP30_CT is the 99th position of SEQIDNO:1, and its base is C / T; the site HdhSNP32_CT is the 138th position of SEQIDNO:2, and its base is C / T; The site HdhSNP36_CA is the 107th position of SEQIDNO:3, and its base is C / ...

Embodiment 2

[0033] This embodiment uses the primer set and probe pair of the above-mentioned SNP site to verify the identification of Chinese and Japanese populations of Abalone wrinkled, including the following steps:

[0034] 1) Genomic DNA extraction of wrinkled abalone: ​​Take about 0.1g of foot muscle, add 500μl STE lysis buffer, cut into pieces, add 50μl 10% SDS, 5μl proteinase K (20mg / ml) in turn, lyse at 56°C for about 3 hours, until the lysate is clear . Add the same volume of Tris saturated phenol (250 μl), chloroform / isoamyl alcohol (24:1) (250 μl), shake gently for 20 minutes, and centrifuge at 12000 rpm for 10 minutes. Take the supernatant and repeat the above steps until there is no protein layer between the aqueous phase and the organic phase. Take the supernatant, add the same volume of chloroform / isoamyl alcohol (24:1), shake gently for 20min, and centrifuge at 12000rpm for 10min. Take the supernatant, add 1 / 10 volume of 3M NaAc (pH5.2) and 2 volumes of cold absolute et...

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Abstract

The invention aims to provide an SNP (single nucleotide polymorphism) marker for distinguishing Chinese population and Japan population of haliotis discus hannai. The SNP marker is SNP loci for distinguishing Chinese population and Japan population of haliotis discus hannai. The invention further provides a primer group and a probe for detecting the SNP loci. The SNP marker is used for identifying the Chinese population and the Japanese population of haliotis discus hannai, so that breeding parent identification at a group level is simpler, more convenient and more feasible.

Description

technical field [0001] The invention belongs to the technical field of aquaculture molecular markers, and in particular relates to a SNP marker for identifying Chinese and Japanese populations of abalone wrinkled. Background technique [0002] Haliotis discus shannai belongs to the phylum Molluscs, Class Gastropoda, and is distributed in Liaoning, Shandong Peninsula, Northeast Japan and other regions in China. It is an important economically cultivated shellfish in northern my country. In the mid-to-late 1990s, in order to reduce the production and survival rate of aquaculture wrinkled abalone brought about by the decline of germplasm, my country began to introduce wrinkled abalone from Japan and hybridize it with local populations to produce hybrid abalone. The hybrid abalone produced by the hybridization of Chinese and Japanese populations has obvious hybrid vigor and has been widely used in aquaculture production. However, in the process of hybrid breeding, it is difficu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 胡晓丽包振民于茜李仰平王师张玲玲陆维
Owner OCEAN UNIV OF CHINA
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