PCR-RFLP identification method for seven crassostrea oyster on south China coast

A technique of PCR-RFLP and giant oyster, applied in the field of molecular biology, achieves the effect of high reliability and wide application range

Active Publication Date: 2008-02-27
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

This method has not been publicly reported in the identification of oyster species in my country, which will provide a scientific method for the identification of species of the genus Pyramid distributed along the coast of South China, and provide technical support for the development of selective breeding and aquaculture of oysters

Method used

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  • PCR-RFLP identification method for seven crassostrea oyster on south China coast
  • PCR-RFLP identification method for seven crassostrea oyster on south China coast
  • PCR-RFLP identification method for seven crassostrea oyster on south China coast

Examples

Experimental program
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Effect test

Embodiment 2

[0052] Example 2 Using COI Fragment PCR-RFLP to Identify the Species of Giant Oyster Ariake

[0053] (1) Genomic DNA extraction

[0054] Take 0.1 g of adductor muscle tissue, add 700ul extraction buffer to digest overnight at 37°C; use an equal volume of phenol-chloroform-isoamyl alcohol (25:24:1) mixture, shake and mix well, 12000g / and centrifuge for 10 minutes; take the supernatant and repeat the previous step once; add an equal volume of chloroform, mix well, and centrifuge at 12000g / min for 5 minutes; take the supernatant and add 2 times the volume of absolute ethanol and 1 / 10 Sodium acetate with a volume of 3M; 12000g / min, centrifuge for 12 minutes, remove the supernatant, and wash with 70% ethanol for 1-2 times; after the ethanol is completely evaporated, dissolve the genomic DNA precipitate in 50ul sterile purified water to obtain genomic DNA. Store at 4°C for later use.

[0055] (2) PCR amplification of COI fragment

[0056] The PCR reaction system is 50ul, includi...

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Abstract

The present invention relates to a PCR-RFLP identification method for 7 kinds of Chinese south inshore oysters. Said identification method includes the following several main steps: making genome DNA extraction, PCR amplification of object fragment, selecting proper incision enzyme combination, utilizing selected incision enzyme to enzyme-cut the PCR product and utilizing agarose gel electrophoresis to make detection.

Description

technical field [0001] The invention relates to a novel molecular biology method, and further relates to a PCR-RFLP identification method for seven species of oysters of the genus Pyramid in the coastal areas of South China. technical background [0002] Oyster is one of the most important economic shellfish at home and abroad. Its meat is delicious, known as "milk in the sea", and it has great nutritional and economic value. Oyster farming is the largest and most widely distributed industry in the shellfish farming industry. The total output and unit area output of oyster farming rank first among all cultured species. There are as many as 6-7 species cultivated in our country. The oysters of the genus Pyramid in South China mainly include Pacific oyster (C.gigas), Portuguese oyster (C.angulata), Hong Kong giant oyster (C.hongkongensis), and Ariake giant oyster. (C. ariakensis), Kumamoto oyster (C. sikamea), C. iredalei and C. iredalei sp. However, because the shell shape ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 喻子牛夏建军
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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