Detection method of turbot FF0901 microsatellite marker by utilizing specific primers

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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Problems solved by technology

[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 sequence alleles are too few, polymorphic The inability to effectively distinguish between different groups or individuals when applied; the Development and characterization of 248 novel microsatellite markers in turbot published by Spain Pardo et al. 248 microsatellite loci were screened out, and primers were designed to provide conditions for the construction of a genetic linkage map, but there was no report on the polymorphism of each primer

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