引物浓度与退火温度不当导致巢式PCR非特异性扩增  

Improper Primer Concentration and Annealing Temperature May Cause Nonspecific Amplication in Nested PCR

作　　者：刘阳[1] 杨淑霞[1] 李敏惠[1] 李丽梅[1] 胡松[1] 刘进[2] 邹强[1] 

机构地区：[1]成都医学院科研实验中心,成都610083 [2]成都医学院药学系

出　　处：《成都医学院学报》2008年第2期111-114,共4页Journal of Chengdu Medical College

基　　金：四川省教育厅资助项目(2006A134);成都医学院基金资助项目(06Z2006-001)

摘　　要：目的探索巢式PCR非特异性扩增的产生原因及消除方法。方法以巢式PCR扩增乙型肝炎病毒(HBV)Sgene为模型,分别使用不同退火温度组合与不同引物浓度组合进行巢式PCR扩增,观察并分析实验结果。结果降低引物浓度或提高退火温度均可消除巢式PCR的非特异性扩增,但提高退火温度有可能影响反应的扩增效率,导致产物量下降甚至得不到产物。结论巢式PCR发生非特异性扩增时,可以提高退火温度或降低引物浓度对反应体系进行优化。Objective To find the mason of causing nonspecific amplication in nested PCR and abate it. Methods To amplify HBV S gene by Nested PCR in different primer concentration and annealing temperature and observe the result. Results Both elevating annealing temperature and decreasing primer concentration can abate nonspecific amplication in nested PCR. But elevating anneal- ing temperature may affect the amplifying effciency that result to decrease or eliminate the PCR products. Conclusion When nonspecific amplication happened in nested PCR, both elevating annealing temperature and decreasing primer concentration can optimize the reaction system.

关 键 词：巢式PCR HBV S GENE 非特异性扩增 

分 类 号：R33[医药卫生—人体生理学]