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Method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence

A cytochrome, mitochondrial technology, applied in the direction of biochemical equipment and methods, determination/inspection of microorganisms, etc.

Inactive Publication Date: 2011-10-05
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

However, this method first needs to design specific primers by itself, and then use cumbersome software to find suitable restriction endonucleases to specifically cut the DNA of the three kinds of fish, and electrophoresis detection can display the final result

Method used

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  • Method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence
  • Method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence
  • Method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence

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Embodiment 1

[0026] Both Japanese mackerel and Australian mackerel in the present invention are collected in Qinglan Port, Wenchang, Hainan.

[0027] A method for identifying Japanese mackerel and Australian mackerel based on the mitochondrial cytochrome b sequence, comprising:

[0028] (1) The total genomic DNA in the muscle tissue of Japanese mackerel and Australian mackerel was extracted by the phenol-chloroform method;

[0029] (2) PCR amplification reaction

[0030] The specific sequences of general primers L14724 and H15915 are: L14724, 5'-GAC TTG AAA AAC CAC CGTTG-3'; and H15915, 5'-CTC CGA TCT CCG GAT TAC AAG AC-3'.

[0031] The reaction system of PCR includes: 100ngDNA template, 0.2mmol / L dNTPs, each primer 1.0umol / L, 4.0mmol / L MgCl 2 , 5.0uL 10×reaction buffer (reaction buffer), 2U Taq polymerase, then add deionized water to a final volume of 50uL;

[0032] The amplification procedure is as follows: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 45 s, annealing a...

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Abstract

The invention relates to a method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence, which comprises the following steps: respectively extracting genome DNA (deoxyribonucleic acid) in muscle tissues of pacific mackerel and slimy mackerel; carrying out PCR (Polymerase Chain Reaction) amplification reaction; sequencing the PCR product, and correcting; and constructing a Cytb base matrix. The method provided by the invention can be used for identifying the pacific mackerel and slimy mackerel in a simple, quick, accurate and efficient way.

Description

technical field [0001] The invention belongs to the technical field of identification of scombroid fish, in particular to a method for identifying Japanese mackerel and Australian mackerel based on mitochondrial cytochrome b sequence. Background technique [0002] Japanese mackerel (S. japonicus) and Australian mackerel (S. australasicus) occupy an important position in China's fishery economy. Their classification is mainly based on traditional classification methods, using "Japanese mackerel with no spots on the abdomen and Australian mackerel with spots" as the identification The most important basis for both. Because the two forms are very similar, and there are often intermediate transition types between the two, it is not easy to distinguish the two by traditional classification methods, which has also led to disputes over the classification status of the two. The indistinguishability between Japanese mackerel and Australian mackerel directly leads to the fact that th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 程起群黄昊郑将臣
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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