Method for identifying two morphological variants of marsupenaeus japonicus

A morphological variation and identification method technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as the inability to correctly identify variant types and the inconspicuous markings on the carapace

Inactive Publication Date: 2011-02-23
XIAMEN UNIV
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Problems solved by technology

At the juvenile stage of Penaeus japonicus and below juvenile stage, due to the inconspicuous carapace markings, the variant type cannot be correctly identified
A

Method used

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  • Method for identifying two morphological variants of marsupenaeus japonicus
  • Method for identifying two morphological variants of marsupenaeus japonicus
  • Method for identifying two morphological variants of marsupenaeus japonicus

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Embodiment Construction

[0040] The present invention is further illustrated below by way of examples.

[0041] 1) According to the known complete mitochondrial genome sequence of Penaeus japonicus (see the US NCBI website http: / / www.ncbi.nlm.nih.gov / nuccore / 63003723), design a pair of primers for amplifying the cytochrome b gene fragment as follows :

[0042] Forward primer: 5'-caaattgttactgggctctttttagct-3';

[0043] Reverse primer: 5'-cagttagcattacgataaatccggt-3';

[0044] 2) Using the conventional phenol-chloroform extraction method to extract the DNA of Penaeus japonicus variant I and variant II;

[0045] 3) Using the DNA obtained in step 2) as a template, carry out a PCR reaction.

[0046] The reaction system is: 10×PCR buffer (Buffer) 2.5μl (containing 20mmol / L Tris-HCl, 100mmol / L KCl, 0.1mmol / L EDTA, 1mmol / L DTT, 0.5% Tween20), 2.5mmol / L MgCl 2 2.5μl, 2.5mmol / L dNTP2.0μl, 50ng / μl template DNA 2μl, 5U / μl TaqDNA polymerase 0.4μl, forward primer and reverse primer each 1.0μl, ddH 2 O to mak...

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Abstract

The invention provides a method for accurately and quickly identifying two morphological variants of marsupenaeus japonicus, which relates to the marsupenaeus japonicus. The method comprises the following steps of: designing and amplifying primers of cytochrome b gene segments of the marsupenaeus japonicus; and extracting the DNA of the variants I and II of the marsupenaeus japonicus, performing polymerase chain reaction (PCR) by taking the DNA as a template, performing enzyme cutting on PCR products and performing electrophoresis identification on an enzyme-cutting products, wherein the variant I of the marsupenaeus japonicus has the enzyme cutting sites of gatatc and produces the two enzyme-cutting segments of 284bp and 299bp by the enzyme cutting; and the variant II of the marsupenaeus japonicus does not have any enzyme-cutting site and still has the strip of 583bp, so that the two variants of the marsupenaeus japonicus can be identified.

Description

technical field [0001] The invention relates to Penaeus japonicus, in particular to a method for identifying two morphological variants of Penaeus japonicus. Background technique [0002] Penaeus japonicus is widely distributed and inhabits in the south of Hokkaido, Japan, the southeast coast of China, Southeast Asia, northern Australia, eastern Africa and the Red Sea. Adult Penaeus japonicus can be clearly divided into two morphological variants according to the markings on the carapace. The distribution of these two morphological variants has obvious regional differences in geographical distribution, and variant I is mainly distributed in Hokkaido, Japan. From the south to the central South China Sea, variant II is mainly distributed in other regions, and both variants appear in the central South China Sea. In the juvenile and below juvenile stage of Penaeus japonicus, due to the inconspicuous carapace markings, the variant type cannot be correctly identified. At present...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 游欣欣董宏标曾凡荣刘洪涛王军苏永全
Owner XIAMEN UNIV
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