Detection method of turbot FF0901 microsatellite marker by utilizing specific primers

A technology of microsatellite marker and detection method, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of few sequence alleles, no primer polymorphism, low polymorphism, etc. To achieve the effect of the simple method

Inactive Publication Date: 2010-05-12
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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[0002] Before the present invention was made, there were similar reports both at home and abroad. Domestically, Isolation and characterization of polymorphic microsatellite loci from ESTlibrary of turbot (Scophthalmus maximus) and cross-species amplification published by Chen Songlin, etc. The detection method of some microsatellite loci in turbot was reported, but the turbot genome library was not constructed by itself, but selected from existing reports; Sun Xiaowen et al. (ConservGenet 2009) published Isolation and characterization of microsatellite markers for turbo (Scophthalmus maximus) by screening

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  • Detection method of turbot FF0901 microsatellite marker by utilizing specific primers

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[0027] The following describes in detail the present invention's DNA molecular genetic marker technology method on the microsatellite core sequence of turbot FF0901 by means of examples in conjunction with the accompanying drawings.

[0028] Firstly, the turbot genomic DNA was extracted and diluted for use; then, using the microsatellite locus sequences contained in the constructed turbot genome library, specific primers were designed at both ends of the sequence; Perform PCR amplification on the genomic DNA of the population or individuals within the population, and detect the PCR products; use the bands that appear in the products to analyze, determine the genotype of each individual, and obtain the genetic polymorphism map of the locus of turbot .

[0029] 1. Extraction of turbot genomic DNA: a. Take 50-100 mg of fresh muscle tissue and put it into a 1.5 mL centrifuge tube. b. Add DNA extraction buffer [0.4mol / L, NaCl, 10mmol / L Tris-HCl (pH 8.0), 2mmol / L EDTA (pH 8.0), 1% ...

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Abstract

The invention discloses a detection method of a turbot FF0901 microsatellite marker by utilizing specific primers, comprising the following steps: firstly extracting turbot genomic DNA; then utilizing a sequence containing a microsatellite in an enrichment library of the turbot microsatellite; designing specific primers at two ends of the sequence as a positive strand 5'-TTACGCTTCGCATCGCTCAT-3', and a negative strand 5'-TGCCAGGCCACCAGACG-3'; carrying out PCR amplification on the turbot genomic DNA; separating the obtained PCR product through modified polyacrylamide gel electrophoresis; analyzing strips generated on the product to obtain a turbot genetic polymorphism map. The method of the invention is suitable for the detection technologies of turbot population genetic marker, genealogy authentication, genetic linkage map construction and the like; the PCR amplification product presents better polymorphism in turbot population detection; and the method of the invention can rapidly acquire a genetic variation map of the turbot FF0901 microsatellite marker, and conveniently, rapidly, accurately and intuitively detect each individual genotype of the turbot.

Description

Technical field: [0001] The invention belongs to turbot DNA molecular genetic marker technology and relates to a microsatellite marker technology method for detecting turbot FF0901. Background technique: [0002] Before the present invention was made, there were similar reports both at home and abroad. Domestically, Isolation and characterization of polymorphic microsatellite loci from ESTlibrary of turbot (Scophthalmus maximus) and cross-species amplification published by Chen Songlin, etc. The detection method of some microsatellite loci in turbot was reported, but the turbot genome library was not constructed by itself, but selected from existing reports; Sun Xiaowen et al. (ConservGenet 2009) published Isolation and characterization of microsatellite markers for turbo (Scophthalmus maximus) by screening SSR-enriched library has reported the construction of a partial genome library of turbot and the screening of microsatellite sequences, but some primers amplified sequenc...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 马爱军侯仕营
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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