Microsatellite Enrichment by Magnetic Beads in Erythroculter ilishaeformis(Bleeker)

According to the hadro-affinity principle of biotin and streptavidin,microsatellite DNAs were isolated from Erythroculter ilishaeformis genome by combining biotin capture method and radioactive labeling hybridization.In this experiment, 613(38.31%)positive colonies were obtained through 1 400 colonies.Sequencing of 288 positive colonies confirmed that 262 of which contained microsatellite sequence clone and 280 microsatellite loci.From these sequences,204 repeat motifs(about 72.86%) were perfect,49 repeat motifs(about 17.5%)were imperfect,27 repeat motifs(about 9.64%)were compound.Two hundred and forty-three pairs of primers were designed with the software Primer 3.0,50 pairs of primers were composed and screened.Results showed that 30 pairs were used successfully to amplify special fragments,and 12 pairs of them were polymorphism within species.