【摘要】 目的为获取滇池水域硅藻18srDNA多态性检验的特异性引物。方法选取文献报道的两对硅藻18srDNA特异性引物A145F/Dia516R及通用性引物18s F/18sR及自行设计的605F/605R和467F/467R硅藻18srDNA引物分别对滇池水域中的硅藻18srDNA进行PCR扩增、克隆和测序检测,并比较其特异性与敏感性。结果引物特异性强弱依次是605F/605R,467F/467R,18s F/18sR,A145F/Dia516R,其敏感性强弱依次是605F/605R,18s F/18sR,467F/467R和A145F/Dia516R。结论引物605F/605R更适合作为滇池水域的硅藻DNA扩增及检验的备选引物。更多还原

【Abstract】 Objective To obtain the specific primers for the detection of diatom 18srDNA polymorphism in Dianchi waters. Methods The primer 605F/605R and 467F/467R of diatom were designed in this study.The diatom specific primer A145 F/Dia516 R and common primer 18sF/18sR were selected from the published literature. Diatom DNAs were extracted from Dianchi water sample,then they were amplified,cloned,sequenced by four pairs of primers respectively,andthe results were compared to determine the their specificity and sensitivity. Results Primers with the strongest specificity were 605F/605R,those with the second and third strongest were 467F/467R and 18sF/18sR, A145F/Dia516R were the last.605F/605R have the strongest sensitivity,too,18 sF/18sR were the second,467F/467R and A145F/Dia516R were the weakest. Conclusion Primer 605F/605R is more suitable as an alternative pair of primers for DNA amplification and examination of diatoms in Dianchi.更多还原