Construction of microsatellite-enriched library and isolation of microsatellite markers in the ridgetail white prawn(Exopalaemon carinicauda)

A microsatellite-enriched library of the ridgetail white prawn(Exopalaemon carinicauda)was constructed using the method of magnetic bead hybridization enrichment.Genomic DNA of ridgetail white prawn was digested with the restriction endonuclease Hae Ⅲ.DNA fragments from 400 to 1 200 bp were recycled and ligated to Hae Ⅲ adaptors.The ligated DNA fragments were hybridized with biotinylated probe(AG)₁₅.Then,enriched DNA fragments were ligated to pMD18-T vector.947 clones were randomly picked and screened by colony PCR.667 clones were positive.184 clones were randomly selected and sequenced from positive clones.150 clones were found to contain 199 microsatellite sequences.Among these microsatellites,79.4% belonged to perfect type,13.6% belonged to imperfect type and 7.0% belonged to compound type.From the primers designed for the 119 microsatellite loci,64 could amplify expected PCR products and 26 were found to be polymorphic.The number of alleles at each locus ranged from 3 to 16.The observed and expected heterozygosities varied from 0.111 to 0.929 and from 0.246 to 0.932,respectively.The PIC value ranged from 0.231 to 0.909.These new loci will be useful in the study of population genetic structure and conservation in this species.