Single nucleotide polymorphism (SNP) marker related to growth speed of basa fish and application of SNP marker

A growth rate, pangasius technology, which is applied in the determination/inspection of microorganisms, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problem of molecular markers to be excavated, etc., and achieve low-cost results

Inactive Publication Date: 2020-06-09
游兆欣
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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 pangasius breeding at this stage still need to be discovered

Method used

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  • Single nucleotide polymorphism (SNP) marker related to growth speed of basa fish and application of SNP marker
  • Single nucleotide polymorphism (SNP) marker related to growth speed of basa fish and application of SNP marker
  • Single nucleotide polymorphism (SNP) marker related to growth speed of basa fish and application of SNP marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Obtaining of SNP markers related to Pangasius growth rate

[0032] 1.1 Acquisition of pangasius populations

[0033] The group used was pangasius hatched on June 15, 2017 in a pangasius farm in Lingshui, Hainan. On August 2, 2017, 60,000 fry were transferred to a 10-acre pond for further breeding. On May 8, 2018, 500 individuals were randomly selected from the pond, 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 Pangasius genomic DNA extraction

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

[0036] (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;

[0037] (2) After the ethanol is basically volatilized, wash with TE buffer solution (10mmol / ml Tris, 1mmol / ml EDTA, SDS 5%, pH=8.0) for ...

Embodiment 2

[0042] Example 2 Sequencing verification and application of SNP markers related to pangasius growth rate

[0043] 2.1 Extract the genomic DNA in the fin rays of pangasius to be tested

[0044] The pangasius to be tested came from the pangasius group in Example 1, and 200 fish were randomly selected again, and the genomic DNA was 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 pangasius to be tested obtained by the aforementioned extraction as a template, the forward primer F: 5'-CTCCTTGTGTGCGTCCTGGTGTGTCTAA-3' (SEQ ID NO: 2) and the reverse primer R: 5'-AAAACATTGCTGTTTGTTATTTAAAA-3'( SEQ ID NO: 3), amplifying the nucleotide fragment where the SNP to be detected is located. 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 polymer...

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Abstract

The invention discloses a basa fish single nucleotide polymorphism (SNP) marker and an application thereof. The nucleotide sequence of the SNP marker is as shown in SEQ ID NO:1, and the 301st base isG or T from the 5'end of the nucleotide sequence shown in SEQ ID NO:1. The SNP marker provided by the invention is closely related to the growth speed of the basa fish, and can be effectively used formolecular marker assisted breeding of basa fish.

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 pangasius, primer pairs and kits for detecting the aforementioned SNP markers, the use of the aforementioned SNP markers, primer pairs, and kits in pangasius breeding, and detection of pangasius method of growth rate. Background technique [0002] Pangasius belongs to Siluriformes and Miscanthus genus, and is an important freshwater aquaculture species in Southeast Asian countries. Pangasius has strong adaptability to the water environment and low requirements for breeding technology. As a growing species of farming, it is currently very popular in the domestic aquatic product market. In 2017, the global production of pangasius was only 1.1 million tons. In 2018, the output reached 2.87 million tons. In 2019, the global pangasius production will exceed 3 million tons. With the small scale of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 游兆欣
Owner 游兆欣
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