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Method for rapidly accurately discriminating crassostrea rivularis and crassostrea gigas

A technology for Pacific oysters and near river oysters, applied in biochemical equipment and methods, microbiological measurement/inspection, etc., can solve the problems of difficult distinction and inaccurate classification of Pacific oysters and near river oysters, and achieve the effect of low cost and simple operation

Inactive Publication Date: 2014-06-18
DALIAN OCEAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, at present, oysters can only be classified by classical taxonomy, that is, by comparing oyster phenotypes, comparative anatomy, and living habits. There is a problem of inaccurate classification, especially it is difficult to distinguish Pacific oysters from near river oysters.

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  • Method for rapidly accurately discriminating crassostrea rivularis and crassostrea gigas
  • Method for rapidly accurately discriminating crassostrea rivularis and crassostrea gigas
  • Method for rapidly accurately discriminating crassostrea rivularis and crassostrea gigas

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Embodiment Construction

[0017] a. Two kinds of oysters were selected (the Pacific oyster culture group was purchased from Tianjin Aquaculture Farm, and the nearby oyster samples were purchased from Hainan Aquaculture Farm), and the adductor muscle of each oyster was placed in EP tubes, and 70% ethanol was added, -20 Store at ℃ for later use; refer to the classic phenol-imitation extraction method of Sambrook et al. to extract the whole genome DNA of oysters from the frozen oyster adductor muscle, and dissolve the DNA samples in deionized sterilized water at a concentration of 100ng / ml. Freeze at -20°C for later use;

[0018] b. Design a primer sequence suitable for the amplification of the target fragment of the two oyster CaM genes, perform PCR amplification on the sample gene obtained in step a, and obtain the PCR product of the target fragment;

[0019] Upstream primer CaM F: 5' ATGGCCGATCAACTCACAG 3'

[0020] Downstream primer CaM R: 5' GCCTTGAGTGGAAGTCGAT 3'

[0021] c. The obtained PCR pro...

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Abstract

The invention discloses a simple-operation low-cost method for rapidly accurately discriminating crassostrea rivularis and crassostrea gigas. The method is performed according to the following steps: extracting gene of oyster samples; performing PCR amplification on oyster sample gene, so as to obtain PCR products of a targeted segment, wherein the PCR products comprise an upstream primer CaMF: 5'-ATGGCCGATCAACTCACAG-3', and a downstream primer CaMR: 5'-GCCTTGAGTGGAAGTCGAT-3'; performing Bln I single enzyme digestion on the obtained PCR products, and performing 1.5% agarose gel electrophoresis on the enzyme digestion products; and according to the size of amplification strips, determining that the gene segment capable of being digested to comprise a segment 227bp and a segment 663bp through single enzyme is the gene segment of crassostrea gigas, and the gene segment which cannot be subjected to Bln I single enzyme digestion is the gene segment of crassostrea rivularis.

Description

technical field [0001] The invention relates to a method for identifying oysters, in particular to a method for quickly and accurately identifying near river oysters and Pacific oysters with simple operation and low cost. Background technique [0002] Oysters are delicious and nutritious, and are the economic shellfish with the largest output in the world. They have also been widely promoted and cultivated in my country. Although there are more than 20 kinds of oysters naturally distributed along the coast of my country, only Pacific oysters and near river oysters are the main cultured species. In the basic research, resource development, protection and utilization of oysters, it is necessary to classify oysters first, and then the shape, phenotype, soft weight, color, shell length, shell height, shell width, and nutrition of this oyster can be classified. Composition and reproduction ability were studied. In recent years, although domestic experts have carried out a large...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2521/301C12Q2565/125
Inventor 仇雪梅于旭蓉常亚青王秀利徐进
Owner DALIAN OCEAN UNIV
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