SNP marker relevant to ammonia endurance capacity of epinephelus coioides and use of SNP marker

A grouper and marking technology, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of molecular markers to be excavated, and achieve the effect of promoting the breeding process

Active Publication Date: 2019-06-07
深圳华大海洋科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Obtainment of SNP markers related to ammonia tolerance of Epinephelus covidii.

[0049] 1.1 The acquisition of the grouper grouper.

[0050] The population used was the fertilized eggs of grouper from a grouper farm in Hainan. The parents were 15 males and 26 females (all F1 generations of wild grouper), 2kg of fertilized eggs It was transferred to Shenzhen Huada Ocean Technology Co., Ltd. Baguang base for incubation. Eight months later, 1,000 individuals with fertilized eggs of Epinephelus nigra were randomly selected for the ammonia tolerance test.

[0051] 1.2 Ammonia tolerance test of Epinephelus slantii.

[0052] The experimental fish had an average body length of 18 cm and an average weight of 78 g. The experimental water is sand-filtered seawater, the water temperature is (21±0.5)℃, the salinity is 26-28, the pH is 7.8-8.0, and the dissolved oxygen is 4.5-5.5mg / L. According to preliminary experiments, this specification of Epinephelus obliquus to ammonia ni...

Embodiment 2

[0062] Example 2: Sequencing verification and application of SNP markers related to the ammonia tolerance of Epinephelus slantii.

[0063] 2.1 Extract the genomic DNA from the fin rays of the grouper.

[0064] The grouper to be tested came from the grouper grouper in Example 1. 300 fish were selected, and the genomic DNA was extracted according to the DNA extraction method described in Example 1.

[0065] 2.2 Amplify nucleotide fragments containing SNP sites.

[0066] Using the genomic DNA of each grouper to be tested obtained by the aforementioned extraction as the template, forward primer F: 5'-CGATGTAATCCAACATTTCCTGA-3' (SEQ ID NO. 2) and reverse primer R: 5'-GATTACAGGGGAAAAACTGCAGAGC- 3'(SEQ ID NO. 3), amplify the nucleotide fragment where the SNP to be tested is located.

[0067] Among them, the PCR reaction system in 25μl is: 50-100ng / μl template DNA 1μl, 10pmol / μl primers F and R each 1μl, 10mmol / L dNTP mix 2.0μl, 5U / μl Taq DNA polymerase 0.125μl, 10×PCR The reaction buffer was...

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Abstract

The invention discloses an SNP marker relevant to ammonia endurance capacity of epinephelus coioides and the use of the SNP marker. The nucleotide sequence of the SNP marker is as shown in SEQ ID NO.1, and the 501st basic group from 5' end in the sequence is G or T. The SNP marker is used for molecular marker auxiliary breeding of the epinephelus coioides. The invention further provides a primer and reagent kit for detecting the SNP marker relevant to the ammonia endurance capacity of epinephelus coioides, and the use of the primer and the reagent kit to breeding of the epinephelus coioides. The invention also provides a method for detecting the ammonia endurance capacity of the epinephelus coioides through the SNP marker, the primer and the reagent kit. Through the adoption of the SNP marker disclosed by the invention, the ammonia endurance capacity of the epinephelus coioides can be detected quickly, efficiently and accurately, and further the SNP marker can be effectively used for molecular marker auxiliary breeding of the epinephelus coioides, so that short-time low-cost high-accuracy breeding of the epinephelus coioides of good varieties can be early realized in an auxiliary manner.

Description

Technical field [0001] The present invention relates to a SNP marker and its use, in particular, to a SNP marker related to the ammonia tolerance of Epinephelus nidus and its use. Background technique [0002] Grouper is a valuable marine 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 contributed to increasing the income of fishermen, meeting the demand for national aquatic products, and optimizing the structure of the aquaculture industry. have important meaning. However, the degradation of germplasm and the lack of good varieties are the main bottlenecks for the sustainable and healthy development of the grouper industry. Therefore, cultivating excellent new species of grouper has become the key to the development of my country's grouper industry. On the other hand, high-density farming models, such as fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
Inventor 游欣欣许腾飞阮志强石琼徐军民
Owner 深圳华大海洋科技有限公司
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