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Chinese prawn B683 microsatellite label detecting technique

A microsatellite marker, the technology of penaeus chinensis, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve problems such as specific genetic markers of microsatellite polymorphism maps of shrimp that have not yet been seen, and achieve the effect of simple method.

Inactive Publication Date: 2004-04-14
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

At present, there are no reports on the construction of Chinese shrimp microsatellite polymorphism map, specific genetic markers and pedigree authentication.

Method used

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  • Chinese prawn B683 microsatellite label detecting technique

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[0015] The following examples describe in detail the present invention's method for molecular genetic markers of the Chinese prawn B683 microsatellite core sequence DNA. First, extract the genomic DNA of Penaeus chinensis and dilute it for later use; then use the microsatellite core sequence in the genome library of Penaeus chinensis to design specific primers at both ends of the sequence; PCR amplification of genomic DNA was carried out, and the PCR products were detected; the bands appearing in the products were analyzed to determine the genotype of each individual, and obtain the genetic polymorphism map of Chinese prawns.

[0016] 1. Extraction of Chinese prawn genomic DNA: Take 100 mg of prawn muscle tissue, cut it into pieces, put it into a 1.5ml centrifuge tube, add 500 μl of TE solution (10mmol / L Tris-CI, 10mmol / L EDTA) with pH 8.0, grind it with Rod grind. Add 25 μl of 10% SDS solution and mix well. Add 4 μl of 20 mg / ml proteinase K, mix well, and digest at 55°C for...

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Abstract

The present invention relates to a detection technique of Chinese prawn B683 microsatellite marker, which includes the following steps: firstly, extracting Chinese prawn genomic DNA; utilizing microsatellite sequence contained in Chinese prawn genomic library to design specific primer at two ends of its sequence; and making PCR amplification of genomic DNA of Chinese prawn; and making the PCR product undergo the process of denaturing polyacrylamide gel electrophoresis separation, and analyzing band produced by said product and obtaining the genetic polymorphism map of Chinese prawn.

Description

Technical field [0001] The invention belongs to the Chinese prawn DNA molecular genetic marker technology, and is a method for detecting the Chinese prawn B683 microsatellite marker technology. Background technique [0002] Before the present invention was made, there were similar research reports both at home and abroad, "Quick Screening of Recombinant Positive Clones Containing Microsatellites in Chinese Prawns by PCR Method" and "Chinese Shrimp Microsatellites" published by Xu Peng et al. Screening of DNA" and other papers have reported the construction of a partial genome library of Chinese prawns and the screening of microsatellite sequences, and evaluated the core sequences of microsatellites according to the standards proposed by Weber (1990), but his research Results were limited to the acquisition of microsatellite sequences for research and no further application development. In 1999, the French Institute of Marine Development (IFREMER) found 10 microsatellite mar...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 孔杰刘萍孟宪红张庆文王伟继费日伟王清印
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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