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Method for detecting SNP marker at EC16 locus of palaemon carinicauda

A technology of white shrimp and a detection method, which is applied in the field of DNA molecular genetic markers of white shrimp, can solve problems such as the lack of SNP markers, and achieve the effect of simple method.

Active Publication Date: 2018-09-25
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Method for detecting SNP marker at EC16 locus of palaemon carinicauda

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Embodiment 1

[0018] The present invention also proposes a method for detecting the SNP marker at the EC16 site of the white shrimp, comprising the following steps:

[0019] 1. First, extract the genomic DNA of individuals in different geographical groups or white shrimp groups and dilute them for later use;

[0020] 2. Using the core sequence containing the EC16 SNP marker in the white shrimp transcriptome library, design specific primers at both ends of the sequence;

[0021] 3. Using the primers to perform PCR amplification on the genomic 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 according to the bands that appear, determine the genotype of each individual, and obtain the genetic polymorphism map of the white shrimp;

[0023] 5. The specific primer sequences are respectively: the forward primer sequence is shown in SEQ ID NO....

Embodiment 2

[0024] Embodiment 2, test method:

[0025] 1. Extraction of genomic DNA from the white shrimp: take 100 mg of the 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) with pH 8.0 , grind with a grinding rod; add 25 μL of 10% SDS solution, mix well; add 4 μL of 20mg / mL proteinase K, mix well, digest at 55°C for 2.5-3h; double-distilled phenol extraction twice, 10min each time, 12000 rpm Centrifuge for 5 min, take the supernatant; extract once with phenol:chloroform (1:1), 10 min, centrifuge at 12000 rpm for 5 min, get the supernatant; extract once with chloroform for 5 min, centrifuge at 12000 rpm for 5 min, get 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 fully dissolving with 500 μL of sterilize...

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Abstract

The invention provides primers for detecting a SNP marker at the EC16 locus of palaemon carinicauda and a method for detecting the SNP marker at the EC16 locus of palaemon carinicauda. The method comprises extracting genomic DNA of the individual within different geographical groups or palaemon carinicauda groups, designing specific primers at both ends of the sequence through the core sequence containing the EC16 SNP marker in a palaemon carinicauda transcriptome library, carrying out PCR amplification on the genomic DNA within different geographical groups or palaemon carinicauda groups, detecting the PCR product, carrying out restriction endonuclease digestion on the PCR product, analyzing the bands, determining the genotype of each individual and acquiring the genetic polymorphism mapof palaemon carinicauda. The primers are mainly used for genetic marking, genealogical identification and genetic map construction of the palaemon carinicauda group.

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 EC16 site of white prawn. 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 white sh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2531/113C12Q2521/313C12Q2565/125
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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