Quick detection method for Scylla paramamosain by microsatellite markers from functional genes

A technology of microsatellite markers and pseudo-cave blue crabs, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of small number of microsatellite markers and limit the development of genetics research, and achieve the amplification of bands clear effect

Inactive Publication Date: 2010-07-28
EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

At present, the number of microsatellite markers available in Scylla cryptae is very small, and there is no report of type I (

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  • Quick detection method for Scylla paramamosain by microsatellite markers from functional genes
  • Quick detection method for Scylla paramamosain by microsatellite markers from functional genes

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[0019] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

[0020] 1. Extraction of Genomic DNA of Scylla pseudocaveus

[0021] Cut a small amount of muscle tissue (approximately 100-150 mg) from Scylla pseudocavena and place it in 500 μl tissue extract (10 mmol / L Tris-Cl, pH 8.0; 100 mmol / L LEDTA, pH 8.0; 100 mmol / L NaCl; 0.5% SDS) Minced, added proteinase K (final concentration 20mg / ml) and RNase A (final concentration 100μg / ml), mixed well, digested at 55°C for 2-3 hours until clear...

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Abstract

The invention relates to a quick detection method for Scylla paramamosain by microsatellite markers from functional genes, which comprises the following steps: (1) extracting genomic DNA of the Scylla paramamosain; (2) acquiring a gene sequence containing microsatellite repeats according to a functional gene sequence of the Scylla paramamosain in a GeneBank database; (3) designing microsatellite marker primers; (4) performing PCR amplification on the genomic DNA of different geographic population or different individuals in the population of the Scylla paramamosain; and (5) detecting a PCR product by polyacrylamide gel electrophoresis. The detection method has the advantages of quickness, accuracy, sensitivity and the like, and can intuitively detect genotypes of different individuals of the Scylla paramamosain so as to quickly acquire a polymorphic map for hereditary variation at functional gene microsatellite loci of the Scylla paramamosain; and the microsatellite markers can be applied to the fields such as hereditary variation analysis of the Scylla paramamosain, germ plasm resource protection and management, genetic linkage map construction and the like.

Description

technical field [0001] The invention belongs to the field of detection of DNA molecular genetic markers of Scylla pseudomae, in particular to a rapid detection method for microsatellite markers of functional gene sources of Scylla pseudomae. Background technique [0002] Microsatellite DNA (Microsatellite DNA) exists in almost all eukaryotic genomes. It is a simple sequence repeat (Simple Sequence Repeats, SSR) composed of 1-6 nucleotides. It is a new type of molecule developed in the past ten years. Marking technology. Microsatellites have the advantages of large number, random distribution, high polymorphism, strong repeatability, co-dominant inheritance, and simple operation, and are widely used in population genetic diversity analysis, germplasm resource protection and management, and genetic linkage maps. Construction and QTL mapping and other fields. [0003] Scholars at home and abroad have developed microsatellite markers in most important aquaculture animals. For...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 马洪雨马凌波马春艳
Owner EAST CHINA SEA FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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