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A detection method for the ec12 SNP marker of the white shrimp

A technology of white shrimp and marking, which is applied in the field of DNA molecular genetic markers of white shrimp, can solve the problem of lack of SNP markers, and achieve the effect of simple method

Active Publication Date: 2021-11-26
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no registered SNP marker in GenBank. The number of SNP markers that can be used for genetic linkage map construction and pedigree identification is still extremely scarce. There is no construction of SNP polymorphism map and specific inheritance of white shrimp at home and abroad Research reports on the application of labeling and other aspects

Method used

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  • A detection method for the ec12 SNP marker of the white shrimp

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Effect test

Embodiment 1

[0018] The detection method of the SNP marker of the EC12 site of the white shrimp: comprising the following steps:

[0019] 1. Firstly extract individual genomic DNA from different geographical groups or white prawn groups and dilute it for later use;

[0020] 2. Using the core sequence containing the EC12 SNP in the white shrimp transcriptome library, design specific primers at both ends of the sequence;

[0021] 3. Then use the primers to perform PCR amplification on the genome DNA of individuals in different geographical groups or groups of white prawns, and detect the PCR products;

[0022] 4. Use the PCR product to carry out restriction endonuclease digestion reaction, analyze the bands that appear, determine the genotype of each individual, and obtain the genetic polymorphism map of the white shrimp.

Embodiment 2

[0023] Embodiment 2, test method:

[0024] 1. Extraction of the genomic DNA of the white shrimp:

[0025] Take 100 mg of white shrimp muscle tissue, cut it into pieces, put it into a 1.5 mL centrifuge tube, add 475 μL of TE solution (10 mmol / L Tris-Cl, 10 mmol / L EDTA) at pH 8.0, grind it with a grinding rod; add 10% SDS Solution 25μL, mix well; add 20mg / mL proteinase K 4μL, mix well, digest at 55℃ for 2.5-3h. Extract twice with double-distilled phenol, 10 min each time, centrifuge at 12,000 rpm for 5 min, and take the supernatant; Lift once for 5 minutes, centrifuge at 12,000 rpm for 5 minutes, and take the supernatant. Add 1 / 25 volume of 5mol / L NaCl solution, mix well, then add twice the volume of -20°C absolute ethanol to precipitate DNA for 15 minutes; pick out DNA, wash with 70% ethanol for tens of minutes, and dry the DNA , after being fully dissolved with 500 μL of sterile water, quantitatively dilute it to a concentration of 50 ng / μL for use.

[0026] 2. Design of S...

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Abstract

The invention provides primers for detecting the SNP marker at the EC12 site of the white shrimp. In addition, the present invention also proposes a method for detecting the SNP marker at the EC12 site of the white shrimp, which includes the following steps: firstly extract the genomic DNA of individuals in different geographical groups or white shrimp groups for future use; Containing the core sequence of the EC12 SNP marker, designing specific primers at both ends of the sequence; then using the primers to perform PCR amplification on the genomic DNA of individuals in different geographical groups or groups of white prawns, and detecting the PCR products; The PCR products were used for restriction endonuclease digestion, and the bands were analyzed to determine the genotype of each individual and obtain the genetic polymorphism map of the white shrimp. The invention can quickly obtain the genetic variation map of the EC12 genetic marker gene locus of the white shrimp, and the obtained result can visually detect the genotype of each individual of the white shrimp, thereby distinguishing homozygous and heterozygous individuals.

Description

technical field [0001] The invention belongs to DNA molecular genetic marker technology of white prawn, and is a SNP marker detection method for the genetic polymorphism of the white prawn at the EC12 site. Background technique [0002] Before the present invention was made, there were only research reports on the microsatellite genetic markers of the white shrimp at home and abroad. In China, Zhu Xiaoyu (2010) reported the versatility of the microsatellite markers of the closely related species Penaeus sinensis to the white shrimp, and 2 general markers were screened out. . Jia Shuwen et al. (2011, 2012) used artificially synthesized biotin-labeled (AG)15 probes and magnetic bead enrichment methods to construct a microsatellite enrichment library for the white shrimp genome, and screened out 26 polymorphic microsatellite markers; and Using 12 microsatellite markers to analyze the genetic diversity of the wild populations of Laizhou Bay, Haizhou Bay and Xiangshan ridgetail ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 李吉涛李健刘萍陈萍
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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