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Blue crab ptssr17 microsatellite DNA marker testing technique

A technology of swimming crab three warts, DNA labeling, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems such as the swimming crab three warts that have not been seen, and achieve the effect of simple method

Active Publication Date: 2008-10-29
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

So far, there have been no reports on microsatellite DNA detection technology in Portunus trituberculatus

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  • Blue crab ptssr17 microsatellite DNA marker testing technique

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Embodiment Construction

[0016] The following examples describe in detail the present invention's DNA molecular genetic marker technology in Portunus trituberculatus ptssr17 microsatellite core sequence. Firstly, the genomic DNA of Portunus trituberculatus was extracted and diluted for later use; then, using the microsatellite core sequence containing ptssr17 in the Genome Library of Portunus trituberculatus, specific primers were designed at both ends of the sequence; The genomic DNA of individuals in Portunus trituberculatus populations was amplified by PCR, and the PCR products were detected by polyacrylamide gel electrophoresis; the bands that appeared in the products were analyzed to determine the genotype of each individual, and the ptssr17 core of Portunus trituberculatus was obtained. Polymorphism map of high genetic variation in sequence regions.

[0017] 1. Extraction of Genomic DNA of Portunus trituberculatus: Take 100mg of muscle tissue, cut it into pieces, put it into a 1.5ml centrifuge t...

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Abstract

The invention relates to a detection technology by ptssr17 microsatellite DNA markers in a blue crab. The technology is characterized in that: first, genome DNA of the blue crab is extracted and diluted to reserve; second, by utilizing a ptssr17 microsatellite core sequence in a genomic library of the blue crab, specificity primers are designed at the two ends of the sequence thereof; third, the genome DNA of different geographic groups of the blue crab or individuals in a blue crab group is processed through the PCR amplification by using the primers, and PCR products are processed through the modified polyacrylamide gel detection; finally, bands which are generated in the products are utilized for analyzing so as to determine the genotype of each individual, thereby obtaining a polymorphic map on the enormous genetic variation of the blue crab in the a ptssr17 core sequence area. The polymorphic map that the ptssr17 genetic mark gene locus of the blue crab shows the enormous genetic variation can be obtained rapidly; the method is simple and convenient; the each individual genotype of the blue crab at the locus can be detected intuitively from the obtained results. The detection technology is mainly used in the genetic marks among the blue crab groups, the genealogical identification, the genetic map construction, etc.

Description

technical field [0001] The invention belongs to microsatellite DNA molecular genetic marker technology of Portunus trituberculatus, in particular to a detection technology of ptssr17 microsatellite DNA marker of Portunus trituberculatus. Background technique [0002] Before the present invention was made, there were relevant research reports in other crabs both at home and abroad. In the study of Dungeness crab (Cancer magister), Pamela C.Jensen et al. designed 99 microsatellite primers, and among them 9 of them were synthesized. In the further study, 6 primers were selected for further study according to the polymorphism of the amplified product, the size of the allele and whether it is easy to count. Using nylon membrane hybridization method, Masatsugu takhano et al. found five variable microsatellite loci in Scylla serrata (Scylla Serrata). David Gopurenko et al screened 5 microsatellite loci in the study of Scylla serrata (Scyllaparamamosain). Using conventional metho...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 刘萍李健宋来鹏高保全
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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