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Yellow crucian carp high-density SNP (single nucleotide polymorphism) marker screening method and application

A screening method and high-density technology, applied in the field of molecular biology, can solve the problems of low purity of target DNA and complicated sequencing procedures, and achieve the effects of reducing library construction costs, consistent amplification efficiency, and simplifying complex genomes

Inactive Publication Date: 2019-03-08
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the sequencing procedure of this invention is relatively complicated, and the purity of the target DNA is low

Method used

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  • Yellow crucian carp high-density SNP (single nucleotide polymorphism) marker screening method and application
  • Yellow crucian carp high-density SNP (single nucleotide polymorphism) marker screening method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0030] The high-density SNP marker screening method of yellow crucian carp comprises the following steps:

[0031] Genomic DNA extraction: Cut the crucian carp muscle or fin ray tissue sample into pieces and place in extraction solution I: 70ml of Tris-HCl and disodium hydrogen phosphate with a concentration of 120mM, 0.3g of fatty alcohol ether ammonium sulfate, and 30ml of chlorine with a concentration of 120mM Sodium chloride solution; add 0.5 μg plasmid DNA and 0.1 μg chitin, freeze and thaw 2-3 times, the freezing temperature is -166°C, the melting temperature is 50°C, centrifuge, collect the supernatant and add extract II: 10ml Tris-HCl with a concentration of 100mM and disodium hydrogen phosphate and 30ml of cationic polyacrylamide with a concentration of 0.5ppm were mixed upside down, centrifuged to produce a white precipitate, and the precipitate was washed to obtain a DNA sample for use; circular DNA and chitin The special existence, on the one hand, can promote the ...

Embodiment 2

[0038] The high-density SNP marker screening method of yellow crucian carp comprises the following steps:

[0039] Genomic DNA extraction: Mince the crucian carp muscle or fin ray tissue sample and place in extraction solution I: 70ml of Tris-HCl and disodium hydrogen phosphate with a concentration of 150mM, 0.5g of fatty alcohol ether ammonium sulfate, and 30ml of chlorine with a concentration of 130mM Sodium chloride solution; add 0.5 μg plasmid DNA and 0.1 μg chitin, freeze and thaw 2-3 times, the freezing temperature is -166°C, the melting temperature is 50°C, centrifuge, collect the supernatant and add extract II: 10ml Tris-HCl with a concentration of 100mM and disodium hydrogen phosphate and 30ml of cationic polyacrylamide with a concentration of 1.6ppm were mixed by inversion, centrifuged to produce a white precipitate, and the precipitate was washed to obtain a DNA sample for use;

[0040] RAD library construction and high-throughput sequencing: the extracted genomic D...

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Abstract

The invention discloses a yellow crucian carp high-density SNP (single nucleotide polymorphism) marker screening method, which includes extracting genomic DNA, constructing a RAD library and high-throughput sequencing, filtering original sequence data, and detecting and screening SNP information sites; adding circular DNA and chitin in the process of extracting the genomic DNA to effectively improve the screening accuracy of SNP markers; and adding chitin fluid into a PCR (Polymerase Chain Reaction) amplification system to improve the reaction sensitivity of PCR amplification and help to obtain the complete high-density SNP sites. The high-density SNP sites obtained by the method can be applied to analyze the genetic diversity level, population genetic structure and the like of the yellowcrucian carp population. The method for obtaining the high-density SNP marker sites of the yellow crucian carp has the advantages of simplicity, high efficiency, high SNP marker screening accuracy, good repeatability, low cost and the like.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a high-density SNP marker screening method and application of yellow crucian carp. technical background [0002] Yellow crucian carp (Ilisha elongate), belonging to Clupeiformes, anchovy family (Engraulidae), and yellow crucian carp (Setipinna), is a warm-water middle-lower economic fish widely distributed in the coastal areas of my country. There is output. Crucian crucian carp mainly feeds on plankton, including diatoms, small swimming animals (juvenile fish), copepods, krill, long-tailed and mysis, etc., and also feeds on benthic organisms and small swimming animals. The prey objects of a variety of high-trophic fish species play an important role in connecting the preceding and the following in the food chain of the marine ecosystem. Its spawning grounds are distributed along the coast and in the inner bay. After spawning every spring, they will ask for bait in var...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869C12Q1/6888
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2531/113C12Q2535/122
Inventor 刘炳舰柳意樊吕振明刘立芹龚理江丽华朱科桦张伟男刘珠段雯
Owner ZHEJIANG OCEAN UNIV
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