Construction and Analysis of Enriched Microsatellite Library of Rhyacionia leptotubula

Objective:To construct an enriched microsatellite library of Rhyacionia leptotubula.Method: Genomic DNA of R.leptotubula was extracted and digested with RsaⅠ.It was hybridized with biotin labeled probe(CT)10 and(GT)10.The hybridized products were enriched with magnetic beads.They were amplified by PCR,and the PCR products were cloned into pMD18-T vector and transferred into Escherichia coli DH5α.Finally,an enriched microsatellite library was constructed.Result: 100 clones were randomly sequenced,and 98 microsatellites were obtained.Among them,the microsatellite clones with repeat motif repeated 5 times or more were accounted for 26%,some of the repeat motifs were with highest repeated times of 33,and 12% of them were imperfect microsatellite,indicating that the constructed library was with high quality.Conclusion: This study was beneficial for isolating polymorphic microsatellites to investigate the population genetic structure,migration,and spreading of R.leptotubula.