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Method for rapid identification of sinonovacula constricta population by utilizing mitochondrial molecular marker

A molecular marker and mitochondrial technology, which is applied in the field of identifying constricted razor clams by using mitochondrial molecular markers, can solve the problems of mixed seedlings and indigenous seedlings, difficult identification, and easy to be affected by environmental factors, and achieves strong reliability and identification methods. simple effect

Inactive Publication Date: 2012-11-14
SHANGHAI OCEAN UNIV
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Problems solved by technology

The introduction and cultivation of off-site seedlings can easily lead to the mixing of off-site seedlings and indigenous seedlings. Therefore, how to distinguish seedlings from different regions for later seed selection, germplasm resource protection and improved seed selection has become an important issue.
Because the phenotypes of different populations of razor clams are similar and are easily affected by environmental factors, it is quite difficult to identify populations based on phenotypes.

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  • Method for rapid identification of sinonovacula constricta population by utilizing mitochondrial molecular marker

Examples

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Embodiment 1

[0024] Example 1: Using the method of the present invention, the Jianghu population and the Zhejiang-Fujian population of razor clam were identified.

[0025] (1) Collect individuals of razor clams from coastal beaches of Jiangsu, Shanghai, Zhejiang and Fujian, and extract DNA;

[0026] The individuals of razor clams were randomly collected from the coastal beaches of Jiangsu, Shanghai, Zhejiang and Fujian. The mantle tissues of razor clam individuals in four regions were collected by scalpel and tweezers, and stored in absolute ethanol for later use.

[0027] DNA was extracted by phenol-chloroform extraction, the specific method is as follows. Take 0.5 g of mantle tissue from each sample and cut it into pieces, add 500 μL of tissue homogenization buffer (10 mmol / dm3 Tris-Hcl, PH=8.0; 50 mmol / dm3 EDTA, PH=8.0), mix well and add to a final concentration of 1 % SDS and 200μg / cm3 of proteinase K, digested and clarified at 55°C. Use saturated phenol to extract once, then use an...

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Abstract

The invention relates to a method for rapid identification of a sinonovacula constricta population by utilizing a mitochondrial molecular marker. The method comprises the following steps of: (1) collecting sinonovacula constricta individuals and extracting DNA (deoxyribonucleic acid); (2) synthesizing a mitochondrial cytochrome oxidase I marker primer; (3) performing PCR (polymerase chain reaction) amplification on genes of the mitochondrial cytochrome oxidase I (COI) of the sinonovacula constricta population; (4) purifying a PCR product; and (5) performing sequencing and sequence alignment on the purified PCR product. The method has the advantages of stable result, strong repeatability and identification simplicity.

Description

technical field [0001] The invention relates to germplasm identification of razor clams, in particular to a method for identifying populations of razor clams by using mitochondrial molecular markers. Background technique [0002] The razor clam is one of the four marine aquaculture shellfish in my country. It has the characteristics of fast growth, short breeding cycle, easy management, high yield and good benefit. According to statistics, the aquaculture output of shellfish in my country accounts for 82% of the total output of mariculture. The annual output of tidal flat shellfish is nearly 2 million tons, of which the output of razor clams has reached 700,000 tons, accounting for about 30% of the total output of tidal flat shellfish aquaculture in China, and occupies an important position in my country's mariculture. [0003] For example, the coastal areas of Zhejiang and Fujian in my country are important seed production areas of razor clams, which are transported to oth...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 牛东红李家乐沈和定王劦金凯
Owner SHANGHAI OCEAN UNIV
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