A Specific Molecular Identification Method of Oyster Omi Omi and Oyster Kumamoto

A technology of Kumamoto oyster and Omi oyster, applied in the field of specific molecular identification of Omi oyster and Kumamoto oyster, can solve the problems of oyster farming, wild resource protection and ecological restoration work in sustainable development and utilization, etc., and achieves low cost and time-consuming. Short, easy-to-use effects

Active Publication Date: 2019-02-15
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is not only a difficult problem in shellfish taxonomy, but also unfavorable for oyster farming, wild resource conservation and sustainable development and utilization, and ecological restoration

Method used

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  • A Specific Molecular Identification Method of Oyster Omi Omi and Oyster Kumamoto
  • A Specific Molecular Identification Method of Oyster Omi Omi and Oyster Kumamoto
  • A Specific Molecular Identification Method of Oyster Omi Omi and Oyster Kumamoto

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Experimental program
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Embodiment 1

[0037] 1. Collection of experimental materials and DNA extraction

[0038] Through morphological identification, the muscle tissues of Oyster Kumamoto and Oyster Omi were collected along the coast of the East China Sea. The total genomic DNA in the muscle tissue of two oysters was extracted by the classic phenol-chloroform method, and stored at -20°C for future use.

[0039] Wherein, the method for extracting total genomic DNA by phenol-chloroform method:

[0040] (1) Treatment before digestion.

[0041] Blot the liquid in the sample with filter paper, take the dorsal tissue and add it to the corresponding centrifuge tube, add 1000 μL of Tris-Cl (pH=8.0) with a final concentration of 1mol / L to the centrifuge tube, and cut it fully with sterilized surgical scissors. Crushed, placed at room temperature for 1h. 4°C, 12000rpm / min, centrifuge for 10min. Remove the supernatant.

[0042] (2) Digestion.

[0043] Add 600 μL of Tris buffer, 50 μL of 10% SDS, and 10 μL of proteinas...

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Abstract

The invention relates to a specific molecule identification method for Cassostrea rivularis and Cassostrea sikamea. The method comprises steps as follows: extracting genomic DNA of the Cassostrea rivularis and the Cassostrea sikamea; performing PCR (polymerase chain reaction) amplification on the genomic DNA by using upstream and downstream primers to obtain PCR products of target fragments, wherein the upstream primer is 5'-GCTTTGGGAGCACTA-3', and the downstream fragment is 5'-CACGAAATTATTCTACATTG-3'; performing gel electrophoresis on the PCR products of the target fragments directly, and judging categories of species according to the size of PCR product strips. The Cassostrea rivularis and the Cassostrea sikamea can be identified economically, quickly and conveniently with the method; the method can play important roles in aquaculture, resource survey and environmental protection.

Description

technical field [0001] The invention belongs to the field of species identification of aquatic organisms, in particular to a method for specific molecular identification of Oyster near the river and Oyster Kumamoto. Background technique [0002] Oyster reef is a kind of biological reef system formed by a large number of oysters gathering and growing on the surface of hard substrate, which is widely distributed in temperate estuaries and coastal areas. Oyster reef restoration is one of the important means to conserve inshore fishery resources and restore the ecosystem of inshore fishery waters. Near River oyster (Crassostrea ariakensis) and Kumamoto oyster (Crassostrea sikamea) coexist in the Yangtze River Estuary, Hangzhou Bay and the coast of Jiangsu, and are the main reef-building oyster species in the East my country Sea. However, it is still unknown which oysters to choose for oyster reef restoration projects in special areas can achieve better ecological restoration ef...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/686C12Q2565/125
Inventor程起群逄娇慧全为民吕浩徐亚岩
OwnerEAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI