Detection method of palaemon carinicauda EC12 SNP marker
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A technology of white shrimp and 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: 2018-11-30
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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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
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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. Using 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 the muscle tissue of white prawns, 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, and 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 12000 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. D...
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Abstract
The invention provides a primer for detecting a palaemon carinicauda EC12 SNP marker. In addition, the invention also provides a detection method of the palaemon carinicauda EC12 SNP marker. The method comprises the following steps of firstly extracting genome DNA of individuals in different geographical groups or palaemon carinicauda groups for use; using core sequences containing EC12 SNP markers in a palaemon carinicauda transcriptome library; designing specific primers at the two ends of the sequence; then, using the primer for performing PCR amplification on the genome DNA of individualsin different geographical groups or palaemon carinicauda groups by using the primer; detecting the PCR products; using the PCR products for detection; performing restriction endonuclease enzyme digestion reaction by using PCR products; performing analysis according to occurring strips; determining the gene type of each individual; obtaining the genetic polymorphism map of the palaemon carinicauda.The genetic polymorphism map of the EC12 genetic marker seat can be fast obtained; the obtained result can be used for directly detecting the gene type of each individual of the palaemon carinicauda,so that the homozygote and heterozygote individuals can be distinguished.
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 ...
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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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