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Aquatic product animal SNP mark screening method

A screening method and technology for aquatic animals, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of low efficiency and high cost

Inactive Publication Date: 2009-01-14
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

[0006] The invention provides a SNP marker screening method for aquatic animals, which can solve the problems of low efficiency and high cost in the prior art

Method used

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  • Aquatic product animal SNP mark screening method
  • Aquatic product animal SNP mark screening method
  • Aquatic product animal SNP mark screening method

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Embodiment 1

[0039] Taking semi-smooth tongue sole as an example below, the technical contents of the present invention are elaborated in conjunction with the accompanying drawings:

[0040] Such as figure 1 As shown, it includes six steps: 1) genomic DNA extraction; 2) MseI restriction endonuclease digestion, adapter ligation and pre-amplification; 3) hybrid duplex formation, CEL I digestion and Bst DNA polymerase Extension; 4) Magnetic bead separation of DNA molecules containing SNP sites; 5) Amplification and enrichment of target DNA and cloning sequencing; 6) Identification of SNP markers. in,

[0041] 1) Genomic DNA extraction

[0042] The DNA of 10 individuals of tongue sole was extracted by phenol-chloroform method. Take 10-20 mg of liver tissue and put it into a mortar, add a little liquid nitrogen and grind until the tissue is ground into powder. Add 1ml of DNA extraction solution TENS (10mM Tris-HCl pH 8.0, 100mM NaCl, 25mM EDTA, 0.5% SDS) for homogenization. Add an equal vo...

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Abstract

The invention provides an aquatic animal SNP marking and selection method, which can solve the problems of low efficiency and high cost existing in the current SNP marking and selection technique. The method adopts the technical proposal comprising the following steps: firstly, genome DNA abstraction; secondly, MseI restrictive enzymes cutting, connection of connecting ends and pre-amplification; thirdly, heterozygous double chain forming, CEL I enzyme cutting and Bst DNA polymerase extending; fourthly, separation of beads containing SNP locus DNA molecule; fifthly, object DNA amplification, concentration and clone sequencing; and sixthly, SNP locus identification. The new SNP marking and selection method can efficiently, plentifully and randomly separate SNP under the condition of unknown genome information, thereby being favorable for the development and the subsequent utilization of the abundant SNP, and having significant application value and popularization prospect in aquatic animal SNP marking and selection, germplasm identification, genetic diversity evaluation, molecular breeding and the like.

Description

technical field [0001] The invention belongs to the screening technology of aquatic animal molecular markers in the technical field of aquatic organisms, and in particular relates to a new method for screening SNP markers of aquatic animals, which is suitable for discovering and screening SNP molecular markers in aquatic animals. Background technique [0002] SNP (Single Nucleotide Polymorphism) as a third-generation molecular marker has the following characteristics: SNP is a biallelic marker; SNP is unevenly distributed on DNA molecules; SNP has a high density; SNP has genetic stability The detection and analysis of SNP can be easily automated. SNP plays an increasingly important role in the construction of high-density genetic linkage maps and the screening of trait-related molecular markers. The acquisition of a large amount of genome sequence information is crucial to the discovery of SNPs, so the current SNP research mainly focuses on humans and various model organism...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 陈松林徐建勇
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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