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Method for microsatellite marker-based genealogy authentication of scylla paramamosain

A technology of microsatellite markers and pseudo-cave blue crabs, which is applied in the measurement/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as microsatellite molecular markers that have not yet been seen, achieve good application prospects, and promote the effect of working health

Inactive Publication Date: 2012-12-12
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Abstract
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Problems solved by technology

At present, there is no research report on the application of microsatellite molecular markers in the selective breeding of Scylla cryptae

Method used

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  • Method for microsatellite marker-based genealogy authentication of scylla paramamosain
  • Method for microsatellite marker-based genealogy authentication of scylla paramamosain
  • Method for microsatellite marker-based genealogy authentication of scylla paramamosain

Examples

Experimental program
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Effect test

Embodiment 1

[0027] 1. Extraction of Genomic DNA from Scylla pseudocaveus

[0028] Cut a small amount of muscle tissue (100-150 mg) from the appendage of Scylla pseudocaveus, and place it in 500 μl of tissue extract (10 mmol / L Tris-Cl, pH8.0; 100 mmol / L EDTA, pH8.0; 100 mmol / L NaCl; 0.5 wt% SDS) was chopped, added proteinase K (final concentration 20mg / ml) and RNaseA (final concentration 100μg / ml), mixed well, digested at 55°C for 2-3 hours until clarification; use phenol: chloroform : Isoamyl alcohol mixture (volume ratio 25:24:1) was extracted twice; then the DNA was precipitated with 2 times the volume of absolute ethanol, washed with 70vol% ethanol solution and dried naturally, then dissolved in 50 μl TE buffer (10mmol / L Tris-HCl, pH8.0; 10mmol / L EDTA, pH8.0); finally the DNA concentration was diluted to 100ng / μl, and stored at -20°C for future use.

[0029] 2. PCR amplification of genomic DNA

[0030] Use 7 pairs of microsatellite primers to carry out PCR amplification of the DNA of...

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Abstract

The invention relates to a method for microsatellite marker-based genealogy authentication of scylla paramamosain. The method comprises the following steps of 1, genomic DNA extraction, 2, microsatellite primer PCR amplification, 3, denaturing polyacrylamide gel electrophoresis of amplification products, and 4, family discrimination. The method provided by the invention can overcome physical marker defects, can realize accurate and effective discrimination of scylla paramamosain individuals from different families, is suitable for identifying mix-bred different individuals from multiple families in family breeding, can promote healthy and fast development of artificial breeding of scylla paramamosain, and has a good application prospect.

Description

technical field [0001] The invention belongs to the field of molecular marker-assisted breeding of mud crabs, and in particular relates to a microsatellite marker-based pedigree authentication method for mud crabs. Background technique [0002] Scylla paramamosain belongs to Arthropoda, Crustacea, Decapoda, Portunidae, and Scylla. It is mainly distributed in the Yangtze River Estuary and beyond in my country. The south coastal waters are my country's important marine fishery resources and seawater cultured crabs. Pseudomonas blue crab has the advantages of large size, fast growth, delicious meat, etc., and is favored by consumers. In recent years, the annual output of artificial breeding of blue crabs in my country has been stable at more than 100,000 tons, maintaining a healthy development momentum. [0003] In the selective breeding of mud crab families, it is necessary to mix the cultivated families in one pond to maintain the consistency of the breeding environment and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 马洪雨马群群马凌波马春艳乔振国
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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