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Method for quickly and accurately identifying northern snakehead, Taiwan snakehead and hybrid snakehead

A technology for hybrid snakehead and snakehead, which is applied to snakehead and hybrid snakehead [snapper (♀) ╳ snakehead (♂)], can quickly and accurately identify the field of snakehead, and can solve the problem of strong subjectivity and inability to effectively identify three species of fish fry, etc. problem, to achieve the effect of accurate identification method

Inactive Publication Date: 2013-02-06
EAST CHINA NORMAL UNIV +2
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for fast and accurate identification of snakeheads, snakeheads and hybrid snakeheads [snakeheads (♀) ╳ snakeheads (♂)] to overcome the existing morphological identification of snakeheads, snakeheads and hybrid snakeheads [ Spotted snakehead (♀)╳black snakehead (♂)], there is a defect of strong subjectivity and cannot effectively identify three kinds of fish fry species, the identification method of the present invention is more accurate and fast

Method used

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  • Method for quickly and accurately identifying northern snakehead, Taiwan snakehead and hybrid snakehead

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Embodiment

[0034] 1. Primer synthesis: According to a new PCR-based molecular marker technology-Related Sequence Amplified Polymorphism proposed by Dr. Li and Quiros from the Department of Vegetable Crops, University of California, USA in 2001, the sequence numbers of the synthetic primers are attached in Table 1;

[0035] Schedule 1

[0036]

[0037] 2. DNA extraction steps of snakeheads, snakeheads and hybrid snakeheads [snakeheads (♀) ╳ snakeheads (♂)]:

[0038] 1) Take a small amount of pectoral fins of snakeheads, snakeheads and hybrid snakeheads [snakeheads (♀)╳ snakeheads (♂), cut them into pieces with ophthalmic scissors and put them into a 1.5ml centrifuge tube;

[0039] 2) Add lysate, which includes 25 μl of Tris-HCl, 50 μl of SDS, 100 μl of EDTA, 320 μl of sterile water, and 5 μl of proteinase K, put it in a 55°C water bath for 2-4 hours, and then it will become clear;

[0040] 3) Add saturated phenol (500 μl) equal to the volume of the lysate, shake well for 10 minutes, a...

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Abstract

The invention relates to a method for quickly and accurately identifying northern snakehead, Taiwan snakehead and hybrid snakehead. The method comprises the steps as follows: 1) synthesizing primers; 2) extracting DNAs (Deoxyribose Nucleic Acids) from the northern snakehead, the Taiwan snakehead and the hybrid snakehead [the northern snakehead (Venus symbol) * the Taiwan snakehead (Mars symbol)]; 3) carrying out SRAP-PCR (Sequence-Related Amplified Polymorphism-Polymerase Chain Reaction) reaction by adopting a renaturation temperature change method; 4) carrying out electrophoresis detection; and 5) combining the primers capable of being used for identifying the northern snakehead, the Taiwan snakehead and the hybrid snakehead [the northern snakehead (Venus symbol) * the Taiwan snakehead (Mars symbol)], processing stripes by using a Gelpro32 software, marking the sizes of the stripes, and identifying in other sampled groups. According to the method, the northern snakehead, the Taiwan snakehead and the hybrid snakehead [the northern snakehead (Venus symbol) * the Taiwan snakehead (Mars symbol)] can be easily identified by utilizing an SRAP technology by cutting away a small amount of fin rays or other fish tissue parts only without killing fishes.

Description

technical field [0001] The present invention relates to a method for identifying hybrid fish species using molecular biology techniques (SRAP). More specifically, the present invention relates to a method for quickly and accurately identifying snakeheads, snakeheads and hybrid snakeheads [snakehead (♀) ╳ snakehead ( ♂)] method. Background technique [0002] Channa argus and Channa maculate belong to the genus Channa in the order Perciformes. The offspring of the hybrid snakehead line obtained through the hybridization of the spotted snakehead and the black snakehead have the advantages of fast growth, high yield, less disease, and high safety and quality. Feed waste and water pollution caused by feeding have become the most important aquaculture species in the Channaidae in the Pearl River Delta. Among them, the hybrid snakehead with the spotted snakehead as the female parent and the black snakehead as the male parent has an increasing proportion in the breeding of snakehe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 朱树人李家乐王群邢智珺谢楠王宇希潘彬斌
Owner EAST CHINA NORMAL UNIV
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