Dasyatis zugei microsatellite sites, primers and application thereof

A technology of microsatellite loci and sharp beaks, which is applied in the field of primers and microsatellite locus sequences of sharp beaked ray, can solve the problems that the molecular genetics research of cartilaginous fishes has not been carried out

Inactive Publication Date: 2014-10-08
张洁
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Problems solved by technology

Its protection requires the understanding of genetic information such as its population structure and geneti

Method used

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  • Dasyatis zugei microsatellite sites, primers and application thereof
  • Dasyatis zugei microsatellite sites, primers and application thereof
  • Dasyatis zugei microsatellite sites, primers and application thereof

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Embodiment Construction

[0012] The present invention will be described in detail below in conjunction with specific embodiments.

[0013] 1. DNA extraction

[0014] Genomic DNA was extracted from the muscle tissue of the sharp-billed ray. SDS / proteinase K cleavage and phenol-chloroform extraction were used for the extraction method (Sambrook et al., 1999). DNA extraction results see figure 1 (display part).

[0015] 2. Construction of Microsatellite Enrichment Library of Stingray Stingray

[0016] Take about 250ng of genomic DNA, use the restriction endonuclease MseI to break, and connect with the AFLP receptor of MseI (5'-TAC TCA GGA CTC AT-3' / 5'-GAC GAT GAG TCC TGA G-3') to connect. After diluting the digestion product tenfold, PCR amplification was performed using AFLP receptor-specific primers (5'-GAT GAG TCC TGA GTA AN-3', referred to as MseI-N). The amplified product was detected using 1% agarose-EB gel, and the result was a diffuse band greater than 200bp, see figure 2 .

[0017] Prob...

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Abstract

The invention discloses dasyatis zugei microsatellite sites, primers and application thereof. The dasyatis zugei microsatellite sites include 10 stable polymorphic microsatellite sites, wherein the sequences of the 10 stable polymorphic microsatellite sites are respectively as shown in SEQ ID NO:1-SEQ ID NO:10. The sequences of the primers of the dasyatis zugei microsatellite sites are respectively as shown in SEQ ID NO:11-SEQ ID NO:30. The application of the dasyatis zugei microsatellite sites or the primers comprises molecular ecology study, pedigree analysis, germplasm resource survey, assistant breeding and individual identification. The genotyping result of 10 pairs of primers in 15 dasyatis zugei individuals shows that the allelic gene number is 3-24, and the average allelic gene number is 13.6; the expected heterozygosity He is 0.5-0.984, and the observed heterozygosity Ho is 0.429-1, so that the 10 pairs of primers are indicated to have high polymorphism and meet the requirement for individual identification of dasyatis zugei.

Description

technical field [0001] The present invention relates, in particular, to a microsatellite site sequence, primer and application thereof. Background technique [0002] The sharp-billed ray (Dasyatis zugei) is a cartilaginous fish that lives in offshore areas, belonging to the order Rayiformes and the family Stingrayidae. Affected by pollution, overfishing, fish farming, etc., the current population is decreasing and needs to be protected urgently. Its protection requires knowledge of genetic information such as its population structure and genetic background. At present, no molecular genetic research on cartilaginous fishes has been carried out in China. Contents of the invention [0003] Aiming at the deficiencies of the prior art, the present invention provides the microsatellite loci and primers of the sharp-billed ray for the first time, and is also the first molecular genetics research result for cartilaginous fishes in China. [0004] Technical scheme of the present ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 张洁陈骁张保卫王慧
Owner 张洁
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