SNP (Single Nucleotide Polymorphisms) marker and application thereof

A technology of labeling and growth rate, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/inspection, etc. It can solve the problems of molecular markers to be excavated and achieve low-cost effects

Active Publication Date: 2017-08-08
ZHENJIANG HUADA DETECTION CO LTD
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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 Polymorphisms) marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Acquisition of SNP markers related to the growth rate of the grouper

[0034] 1.1 Acquisition of the oblique-banded grouper population

[0035] The group used was a grouper hatched on November 10, 2010 in a grouper farm in Hainan. The parents were 29 wild males and 12 wild females (caught in the South China Sea in Sanya, Hainan), in 2010 On December 10, 2009, 15,000 fry were transferred to a cage for further rearing. On August 8, 2011, 198 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.

[0036] 1.2 Genomic DNA extraction of the grouper

[0037] 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:

[0038] (1) Take 0.3-0.5g of fin rays in a 1.5ml Eppendorf tube, cut it into pieces, and dry it for 20 minutes on a clean bench;

[0039]...

Embodiment 2

[0044] Example 2 Sequencing verification and application of SNP markers related to the growth rate of the grouper

[0045] 2.1 Extract the genomic DNA from the fin rays of the to-be-tested grouper

[0046] The grouper to be tested comes from the grouper population in Example 1, 180 fish are randomly selected, and the genomic DNA is extracted according to the DNA extraction method described in Example 1.

[0047] 2.2 Amplification of nucleotide fragments containing SNP sites

[0048] Using the genomic DNA of each tested grouper obtained from the aforementioned extraction as a template, the forward primer F: 5'-CTCCTCTGCTGCTCAGCTTT-3' (SEQ ID NO: 2) and the reverse primer R: 5'-CCGTACCTCCACTGATGTGA- 3' (SEQ ID NO: 3), the nucleotide fragment where the SNP marker 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 0.1...

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Abstract

The invention discloses an SNP (Single Nucleotide Polymorphisms) marker and application thereof. The SNP marker is a 501st basic group C or T from a 5' end of a nucleotide sequence shown as SEQ ID NO: 1. The SNP marker provided by the invention is closely related to a growth speed of saddletail grouper and can be effectively used for molecular marker assisted breeding of the saddletail grouper.

Description

technical field [0001] The present invention relates to SNP markers and their applications. Specifically, the present invention relates to SNP markers related to the growth rate of the grouper, primer pairs and kits for detecting the aforementioned SNP markers, and the aforementioned SNP markers, primer pairs, and kits are used in the breeding of the grouper The use of and the method for detecting the growth rate of the grouper. 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 bottleneck...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 游欣欣王敏马兴宇石琼徐军民
Owner ZHENJIANG HUADA DETECTION CO LTD
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