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SNP (single nucleotide polymorphism) marker and application thereof

A technology of marking and body weight, applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc., can solve the problems of molecular markers that need to be mined

Active Publication Date: 2015-03-18
SHENZHEN HUADA GENE INST +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, molecular markers related to growth traits that can be effectively used in grouper breeding at this stage still need to be discovered

Method used

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  • SNP (single nucleotide polymorphism) marker and application thereof
  • SNP (single nucleotide polymorphism) marker and application thereof
  • SNP (single nucleotide polymorphism) marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Obtaining SNP markers related to the weight of the grouper

[0032] 1.1 Acquisition of the oblique-banded grouper population

[0033] The group used was the grouper hatched on November 10, 2010 in a grouper farm in Hainan. On December 10, 2010, 15,000 fry were transferred to a cage for further rearing. On August 8, 2011, 199 individuals were randomly selected from the cage, and the dorsal fin rays of the fish were cut and stored in 95% ethanol at -20°C for genomic DNA extraction.

[0034] 1.2 Genomic DNA extraction of the grouper

[0035] In this experiment, the conventional phenol-chloroform method was used to extract the genomic DNA in the fin rays of the grouper, and the specific steps were as follows:

[0036] (1) Take 0.3-0.5g of fin rays in a 1.5ml Eppendorf tube, cut into pieces, and dry for 20 minutes on a clean bench;

[0037] (2) After the ethanol is basically volatilized, wash with TE buffer solution (10mmol / ml Tris, 1mmol / ml EDTA, SDS5%, pH=8.0)...

Embodiment 2

[0042] Example 2 Sequencing verification and application of SNP markers related to weight of grouper

[0043] 2.1 Extract the genomic DNA from the fin rays of the to-be-tested grouper

[0044] The grouper to be tested comes from the grouper population in Example 1, 50 fish are randomly selected, and the genomic DNA is extracted according to the DNA extraction method described in Example 1.

[0045] 2.2 Amplification of nucleotide fragments containing SNP sites

[0046] Using the genomic DNA of each tested grouper obtained from the aforementioned extraction as a template, the forward primer F: 5'-ACAGAGGAACAGACTGCGACAC-3' (SEQ ID NO: 2) and the reverse primer R: 5'-RCTGGCAACTAGCAGACGTAGAA- 3' (SEQ ID NO: 3), the nucleotide fragment where the SNP to be detected is located is amplified. Among them, the PCR reaction system is calculated as 25 μl: 50-100ng / μl template DNA 1μl, 10pmol / μl primer F and R 1μl each, 10mmol / L dNTP mix 2.0μl, 5U / μl Taq DNA polymerase 0.125μl, 10×PCR rea...

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Abstract

The invention discloses an SNP (single nucleotide polymorphism) marker and application thereof. The SNP marker is the 381st base T or C from the 5' terminal of a nucleotide sequence as shown in the SEQ ID NO:1. The SNP marker is closely related to the weight trait of epinephelus coioides, and can be effectively used for molecular mark assisted breeding of epinephelus coioides.

Description

technical field [0001] The present invention relates to SNP markers and their applications. Specifically, the present invention relates to SNP markers related to body weight traits of grouper, primer pairs and kits for detecting the SNP markers described in claim 1, the aforementioned SNP markers, primer pairs, and kits are used in grouper Uses in fish breeding, and methods for detecting body weight traits in groupers. Background technique [0002] Grouper is a rare seawater economic fish. In the past ten years, the artificial breeding technology of grouper has been relatively mature, and the scale of grouper farming has gradually expanded. The rapid development of the grouper industry has played an important role in increasing fishermen's income, meeting the national demand for aquatic products, and optimizing the structure of aquaculture industry. have important meaning. However, the degradation of germplasm and the lack of good species are the main bottlenecks for the ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 游欣欣石琼张勇杨成业阮志强束礼平蒙子宁张海发林浩然
Owner SHENZHEN HUADA GENE INST
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