Identification method of channel catfish species

A technology for channel catfish and identification methods, which is applied in the direction of microbial determination/inspection, biochemical equipment and methods, and can solve problems such as cumbersome operation process, lack of mitochondrial DNA molecular data, and unsuitability for rapid and economical identification of channel catfish species. , to achieve the effect of short time and simple operation

Inactive Publication Date: 2013-11-27
罗志萍
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Problems solved by technology

The results of relevant scientific and technological novelty searches show that there are reports abroad on the PCR-RFLP data of mitochondrial DNA for species identification in cod, flounder and other products, but there is no molecular data of the D-loop region of mitochondrial DNA for channel catfish species identification. report
Moreover, the PCR-RFLP method is cumbersome to operate, and it is mostly used for genetic polymorphism analysis of different populations of the same species, which is not suitable for rapid and economical identification of channel catfish species.

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  • Identification method of channel catfish species
  • Identification method of channel catfish species
  • Identification method of channel catfish species

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Embodiment Construction

[0042] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0043] 1. Genomic DNA extraction

[0044] Using the conventional phenol-chloroform extraction method, refer to the method of Sambrook (2002) with slight modifications:

[0045] (1) Take about 0.1 g of the caudal fin fixed with alcohol, put it in a 2ml centrifuge tube, cut it into pieces, and dry it at 37°C;

[0046] (2) Add 500ul of extraction buffer to the centrifuge tube, and add RNaseA to a final concentration of 20μg / ml, and bathe in water at 37°C for 1h. Then add proteinase K to the extraction buffer to a final concentration of 200 μg / ml, and bathe overnight at 55°C;

[0047] (3) After cooling the digested sample to room temperature, add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1), gently...

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Abstract

The invention relates to an identification method of channel catfish species. The identification method comprises following steps: genome DNA of channel catfish muscular tissues is extracted; a PCR primer is designed in mitochondrial DNA D-loop region; the genome DNA is subjected to PCR amplification with the presence of the PCR primer so as to obtain mitochondrial DNA D-loop sequences; the mitochondrial DNA D-loop sequences are subjected to enzyme digestion for 2 to 3h at a temperature of 37 DEG C by using restriction enzyme EcoR I; and then the mitochondrial DNA D-loop sequences are subjected to agarose gel electrophoresis after enzyme digestion, and the channel catfish species are identified according to the results of electrophoresis. Enzyme sites are searched based on sequencing of D-loop, and enzyme digestion identification is performed, so that sequencing is not necessary for identification of channel catfish species. The identification method is capable of identifying channel catfish species at a molecular level; operation is simple and easy; consumed time is short; and identification method is suitable for species identification of channel catfish in import and export trade.

Description

technical field [0001] The invention relates to a method for identifying channel catfish species, in particular to a method for identifying genetic information of channel catfish species by using a mitochondrial DNA D-loop control region, and belongs to the field of molecular biological PCR genetic identification. Background technique [0002] Channel catfish (Ictalurus punctatus), also known as Channel Catfish, belongs to Siluriformes, Ictaluridae, and Ictalurus. It is native to North America, and its natural range is in the central United States, southern Canada, and northern Mexico. Channel catfish is a rare and valuable freshwater economic fish, and is deeply loved by consumers in the United States, Canada and many other countries. Processed finished products or semi-finished products are very popular in the markets of the United States, Japan, Europe and Canada, and the market demand is huge. Since 1984, my country’s Hubei Provincial Fisheries Research Institute intro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 罗志萍王华全肖武汉
Owner 罗志萍
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