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Recombination activating gene (RAG) 2 molecular marker restriction fragment length polymorphism (RFLP) identifying method for aquaculture of catfish hybrids

A technology of RAG2 and aquaculture, which is applied in the measurement/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of low detection efficiency, low repeatability, and influence on popularization and use, and achieve simple operation, fast experiment, and identification intuitive effect

Inactive Publication Date: 2013-09-18
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the RAPD marker used in its identification work is limited by the method itself, and the detection efficiency is low and the repeatability is not high, which seriously affects the popularization and use of this method.

Method used

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  • Recombination activating gene (RAG) 2 molecular marker restriction fragment length polymorphism (RFLP) identifying method for aquaculture of catfish hybrids
  • Recombination activating gene (RAG) 2 molecular marker restriction fragment length polymorphism (RFLP) identifying method for aquaculture of catfish hybrids

Examples

Experimental program
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Effect test

Embodiment 1

[0035] RAG2 molecular marker RFLP identification method for aquaculture catfish hybrids, with southern catfish, catfish and their hybrid F 1 Generation as the research object, including the following steps:

[0036] In the first step, samples of southern catfish, catfish and their hybrid offspring were taken respectively, and genomic DNA was extracted.

[0037]Because the individuals of the hybrid fry are small, the genomic DNA of the fry sample was extracted with the Marine Tissue DNA Extraction Kit (Tiangen Company).

[0038] Genomic DNA was extracted from parental catfish and catfish ray samples by conventional phenol-chloroform extraction (Sambrook & Russell, 2001):

[0039] a. Material collection: Take a small amount of fin ray sample soaked in absolute ethanol, cut it into pieces with alcohol-sterilized scissors, and put it into a 1.5 ml EP tube.

[0040] b Digestion: Add 500 μL lysis buffer (10 mmol / L Tris-Cl; 0.1 mol / L EDTA; 0.5 % SDS; pH 8.0), 5 μL proteinase K (20 ...

Embodiment 2

[0093] RAG2 molecular marker RFLP identification method for aquaculture catfish hybrids, with southern catfish, catfish and their hybrid F 1 Generation as research object, technical scheme is identical with embodiment 1.

[0094] On the basis of Example 1, in the third step, use a PCR instrument to carry out enzyme digestion, and the reaction parameters are respectively:

[0095] xho Ⅰ The parameters for digesting RAG2 gene fragments are: 37°C for 5 minutes, 80°C for 5 minutes.

[0096] StuⅠ The parameters for digestion of the RAG2 gene fragment are: 37°C for 5 minutes, 80°C for 10 minutes.

Embodiment 3

[0098] RAG2 molecular marker RFLP identification method for aquaculture catfish hybrids, with southern catfish, catfish and their hybrid F 1 Generation as research object, technical scheme is identical with embodiment 1.

[0099] On the basis of Example 1, a PCR instrument was used to carry out enzyme digestion, and the reaction parameters were respectively:

[0100] xho Ⅰ The parameters for digesting RAG2 gene fragments are: 37°C for 5 min 30 s, 80°C for 5 min 30 s.

[0101] Stu Ⅰ The parameters for digesting RAG2 gene fragments are: 37°C for 5 min 30s, 80°C for 10 min 30s.

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Abstract

The invention relates to an authenticating method of fish hybrids and particularly relates to an RAG2 molecular marker RFLP identifying method for aquaculture of catfish hybrids. Southern catfishes, catfishes and hybrids of the southern catfishes and the catfishes serve as objects of study, RAG2 gene sections in catfishes are selected to serve as molecular markers, the RFLP analysis of the RAG2 gene sections is achieved, and the southern catfishes, the catfishes and hybrids of the southern catfishes and the catfishes are identified successfully. The method has the advantages of being fast in experiment, convenient to operate, direct in identification and free from sequencing.

Description

technical field [0001] The invention relates to a method for identifying fish hybrids, in particular to a method for identifying RAG2 molecular marker RFLP of aquaculture catfish hybrids. Background technique [0002] Siluriformes are carnivorous demersal fishes and are one of the most developed teleosts in fish evolution. Southern catfish (also known as big mouth catfish) ( Silurus meridionalis Chen) and catfish ( Silurus asotus Linnaeus) belong to Siluriformes, Siluridae, and Silurus genus, both of which are delicious and have high nutritional value, and are high-value-added economic fish mainly cultured in various parts of my country. Southern catfish is distributed in the Pearl River, Minjiang River, Xiangjiang River, Yangtze River and other water systems in my country. It is a large and ferocious carnivorous fish. It has the characteristics of fast growth, large body, and strong disease resistance. It likes to inhabit open water bodies, and The gravel section of the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 宋昭彬郭睿杨世勇张修月
Owner SICHUAN UNIV
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