OBJECTIVE: To analyze the sequence of microsatellite and the flanking sequence from four populations of Oncomelania hupensis. METHODS: We cloned 159 SSR-PCR amplification products of a commonly used primer, (CA)(8RY), using O. hupensis genomic DNA as template, and sequenced 82 products. RESULTS: The sequences obtained were novel O. hupensis genomic sequences but not repeat simple sequence. It was observed that 36 out of 82 clones contained microsatellites between priming sites. The flanking sequences of certain microsatellite were invariant. Both (GA/CT)(n) and (TTAGGG/CCCTAA)(n) were found in four populations of O. hupensis. However, (CAA)(n) were found only in O. hupensis from Fuqing, Fujian province and (TCTCTG)(n) were found only in O. hupensis from Guichi, Anhui province and (GAA/TTC)(n), (CAA/TTG)(n), (CAT)(n) were found only in O. hupensis from Puge, Sichuan province. CONCLUSION: The results obtained by SSR-PCR should not be interpreted as the amplification of microsatellite loci, and analytical rules similar to those for Random Amplified Polymorphic DNA should be used. SSR-PCR could not make the most of the priority of microsatellite. It seems better to amplify the microsatellites with the primers designed on the basis of the flanking sequence.
OBJECTIVE: To analyze the sequence of microsatellite and the flanking sequence from four populations of Oncomelania hupensis. METHODS: We cloned 159 SSR-PCR amplification products of a commonly used primer, (CA)(8RY), using O. hupensis genomic DNA as template, and sequenced 82 products. RESULTS: The sequences obtained were novel O. hupensis genomic sequences but not repeat simple sequence. It was observed that 36 out of 82 clones contained microsatellites between priming sites. The flanking sequences of certain microsatellite were invariant. Both (GA/CT)(n) and (TTAGGG/CCCTAA)(n) were found in four populations of O. hupensis. However, (CAA)(n) were found only in O. hupensis from Fuqing, Fujian province and (TCTCTG)(n) were found only in O. hupensis from Guichi, Anhui province and (GAA/TTC)(n), (CAA/TTG)(n), (CAT)(n) were found only in O. hupensis from Puge, Sichuan province. CONCLUSION: The results obtained by SSR-PCR should not be interpreted as the amplification of microsatellite loci, and analytical rules similar to those for Random Amplified Polymorphic DNA should be used. SSR-PCR could not make the most of the priority of microsatellite. It seems better to amplify the microsatellites with the primers designed on the basis of the flanking sequence.