Primer and method for identifying pseudosciaena crocea of daiqu species and min-yuedong species

A technology of large yellow croaker and Daiqu family, which is applied in the field of biotechnology species identification, can solve the problems of unstable RAPD technology system and low repeatability, and achieve the effects of fast identification, low false positive rate and good repeatability

Active Publication Date: 2012-11-14
NINGBO UNIV
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AI Technical Summary

Problems solved by technology

[0003] The existing Chinese patent name is the identification primers and identification methods of different populations of large yellow croakers (application number is CN200610135259.2) discloses the identification primers and identification methods of large yellow croakers of the Daiqu, Fujian, Guangdong, and Naozhou groups. The RAPD-PCR amplification pattern of the primers can distinguish the large yellow croaker of the Daiqu family and the large yellow croaker of the Fujian-Yuedong family. The technology itself has the disadvantages of unstable system and low repeatability

Method used

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  • Primer and method for identifying pseudosciaena crocea of daiqu species and min-yuedong species
  • Primer and method for identifying pseudosciaena crocea of daiqu species and min-yuedong species
  • Primer and method for identifying pseudosciaena crocea of daiqu species and min-yuedong species

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specific Embodiment 2

[0041] A kind of identification method of Daiqu large yellow croaker and Fujian-Yuedong large yellow croaker of the present invention, concrete steps are as follows:

[0042] (1) Extraction and quantitative detection of total DNA of large yellow croaker

[0043] (2) Preparation of PCR amplification system

[0044] The preparation of the PCR amplification reaction system is as follows: various components and final concentrations are: buffer 10mM, dNTP 0.2mM, Mg 2+ 2mM, each primer is 0.2mM, Taq enzyme 1U, large yellow croaker template DNA 10-50ng, make up to 25ul with double distilled water, the primer sequence is:

[0045] J1 upstream primer 5′-GGCATTTAGGCACAAGGAC-3′

[0046] J1 downstream primer 5′-GGGGGCACAACTCAGAGG-3′

[0047] J2 upstream primer 5′-CAGGCCCCACCATACTAGT-3′

[0048] J2 Downstream Primer 5′-GGGTGCCGGTTCCTGAC-3′

[0049] (3) PCR amplification reaction

[0050] Heat on the PCR instrument at 95°C for 2-5min to fully denature the template DNA, then enter the...

specific Embodiment 3

[0053] A kind of identification method of Daiqu large yellow croaker and Fujian-Yuedong large yellow croaker of the present invention, concrete steps are as follows:

[0054] (1) DNA extraction and quantitative detection of large yellow croaker by phenol-chloroform extraction method

[0055] (2) Preparation of PCR amplification system

[0056] The preparation of the PCR amplification reaction system is as follows: various components and final concentrations are: buffer 10mM, dNTP 0.2mM, Mg 2+ 2mM, each primer is 0.2mM, Taq enzyme 1U, 10ng of large yellow croaker template DNA in the PCR amplification reaction system of primer pair J1, 40ng of large yellow croaker DNA in the PCR amplification reaction system of primer pair J2, filled up with double distilled water to 25ul, the primer sequence is

[0057] J1 upstream primer 5′-GGCATTTAGGCACAAGGAC-3′

[0058] J1 downstream primer 5′-GGGGGCACAACTCAGAGG-3′

[0059] J2 upstream primer 5′-CAGGCCCCACCATACTAGT-3′

[0060] J2 Downs...

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Abstract

The invention discloses a primer and a method for identifying pseudosciaena crocea of daiqu species and min-yuedong species. The primer is characterized in that primer sequences are a primer pair J1 of 5-GGCATTTAGGCACAAGGAC-3 and 5-GGGGGCACAACTCAGAGG-3 and a primer J2 of 5-CAGGCCCCACCATACTAGT-3 and 5-GGGTGCCGGTTCCTGAC-3. The method for identifying specifically comprises the following steps: extracting pseudosciaena crocea total deoxyribose nucleic acid (DNA) and quantitatively detecting; preparing a polymerase chain reaction (PCR) amplification system and carrying out amplification reaction; finally, carrying out agarose gel electrophoresis detection on a product, if bands within the ranges of 300-400bp and 600-700bp of a map appear, determining that the DNA sample is obtained from pseudosciaena crocea of daiqu species; and if not, determining that the DNA sample is obtained from pseudosciaena crocea of min-yuedong species. The method has the advantages of high stability, good repeatability, quick identification speed, low false positive rate and the like.

Description

technical field [0001] The invention relates to the field of biotechnology species identification, in particular to an identification primer and an identification method for Daiqu large yellow croaker and Fujian-Yuedong large yellow croaker. Background technique [0002] Large yellow croaker ( Pseudosciaena crocea ), also known as yellow croaker and yellow croaker, belongs to Perciformes, Sciaenidae, and Pseudosciaena. Largest fish species. The coastal large yellow croaker in my country can be roughly divided into three geographical populations: Daiqu, Fujian-Yuedong and Naozhou. Because of the advantages of the large yellow croaker of the Daiqu family in terms of body color, body shape and flavor, the price in the market is higher than that of the large yellow croaker of the Fujian-Yuedong family; The large yellow croaker of the Daiqu family is lighter in body color; in terms of body shape, the large yellow croaker of the Daiqu family is slender, while the large yellow c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 严小军沈锡权王梦林闫旭红
Owner NINGBO UNIV
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