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Method for detecting ES15 microsatellite markers of Chinese shrimps

A technology of microsatellite marker and Chinese prawn, which is applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., to achieve the effect of simple method

Active Publication Date: 2010-11-10
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 and the application of specific genetic markers at home and abroad.

Method used

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  • Method for detecting ES15 microsatellite markers of Chinese shrimps
  • Method for detecting ES15 microsatellite markers of Chinese shrimps
  • Method for detecting ES15 microsatellite markers of Chinese shrimps

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Embodiment Construction

[0013] In the following, the present invention will be described in detail with reference to the accompanying drawings, and the method for molecular genetic markers of the Chinese prawn ES 15 microsatellite core sequence DNA.

[0014] Firstly, the genomic DNA of Penaeus chinensis was extracted and diluted for later use; then, using the microsatellite core sequence contained in the cDNA library of Penaeus chinensis, specific primers were designed 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.

[0015] 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 475 μl of TE solution (10mmol / L Tris-Cl, 10mmol / L EDTA) with a pH of 8.0, grind with Rod grind. Add 25 μl of 10% SDS so...

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Abstract

The invention relates to a method for detecting ES15 microsatellite markers of Chinese shrimps. The method comprises the following steps of: extracting genome DNA of the Chinese shrimps first; designing specific primers at both ends of a sequence of a cDNA library by utilizing the microsatellite core sequence in the cDNA library of the Chinese shrimps; performing PCR amplification on the genome DNA of individuals in different geographic groups of the Chinese shrimps or Chinese shrimp groups, and performing denaturing polyacrylamide gel electrophoretic separation on obtained PCR products to obtain a high-genetic variation polymorphism map presented by the Chinese shrimps in an ES15 core sequence area; and analyzing stripes of the map to obtain a genotype of each individual of the Chinese shrimps in ES15 loci. Through the method, a hereditary variation map of the ES15 loci of the Chinese shrimps can be obtained quickly, the genotype of each individual of the Chinese shrimps can be detected visually, and the method is simple and convenient; and the method is mainly applied to group genetic markers, genealogical authentication, the construction of genetic maps and the like of the Chinese shrimp.

Description

Technical field: [0001] The invention belongs to the Chinese prawn DNA molecular genetic marker technology, and is a microsatellite marker detection method for the genetic polymorphism of the Chinese prawn at the ES15 site. Background technique: [0002] Before the present invention is made, there are similar research reports both at home and abroad, and domestic Chinese Academy of Sciences Institute of Oceanology Xu Peng etc. (Acta Fisheries, 2001a; Ocean and Limnology, 2001b; Recombination Positive Clones Containing Microsatellites", "Screening of Microsatellite DNA from Penaeus Chinese Shrimp" and "Study on Screening Microsatellite Markers from ESTs of Penaeus Chinese" have reported the construction of a partial genome library of Penaeus Chinese, containing microsatellite sequences According to the criteria proposed by Weber (USA, Marshfield Medical Research Foundation, 1990), the microsatellite core sequences were evaluated, but his research results were limited to the a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 李吉涛李健陈萍宋春妮刘萍王清印
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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