A Molecular Method for Distinguishing Oyster Omi and Oyster Kumamoto

A technology for Omi oysters and Kumamoto oysters, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of oyster farming, wild resource protection and ecological restoration work in sustainable development and utilization, etc., and achieves high accuracy. Effect

Active Publication Date: 2019-03-26
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 Molecular Method for Distinguishing Oyster Omi and Oyster Kumamoto
  • A Molecular Method for Distinguishing Oyster Omi and Oyster Kumamoto
  • A Molecular Method for Distinguishing Oyster Omi and Oyster Kumamoto

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Embodiment 1

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

[0038] Muscle tissues of Oyster Kumamoto and Oyster Omi were collected through morphological identification. 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 proteinase K to the centrifuge tube, shake and mix o...

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Abstract

The invention relates to a method of molecularly identifying crassostrea rivularis and Crassostrea sikamea. The method includes: extracting genome DNA of crassostrea rivularis and crassostrea sikamea; utilizing an upstream primer and a downstream primer to perform PCR (polymerase chain reaction) amplification on the genome DNA to obtain a PCR product of a target segment, wherein the upstream primer is 5'-GATTATTATGCTGGGATTAG-3, and the downstream primer is 5'-ACCCTCTTTCGCTTCA-3; directly performing gel electrophoresis on the PCR product of the target segment, and judging species type according to stripe size of the PCR product. By using the method, crassostrea rivularis and crassostrea sikamea can be identified economically, quickly, simply and conveniently, and results of sequencing verification show that identifying accuracy is 100%. The method is about to play an important role in aquaculture, resource investigation and environment protection.

Description

technical field [0001] The invention belongs to the field of identification of aquatic product species, in particular to a method for molecular identification of Oyster near 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 effects. The ide...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888
Inventor程起群全为民逄娇慧徐亚岩吕浩
OwnerEAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI