摘要： DMSO通常经验性地用于提高PCR扩增的效率,但是过量的DMSO可以显著降低特异序列的扩增效率,尤其是导致非特异性扩增的现象却被忽视.在6%的DMSO存在时,开始出现非特异条带,同时特异扩增产物减少.本文首次报道DMSO对PCR产物特异性的影响并确定了克服上述现象产生的途径,即通过增加模板-引物的比例消除非特异条带.而通常提高复性温度只能部分减少非特异产物,不能避免非特异扩增.本文结果有助于提高常规PCR,尤其是分子遗传学分析中经常使用的随机扩增多态性DNA(randomamplifiedpolymorphicDNA,RAPD)检测的准确度。
Abstract:DMSO is the most often empirically used to increase efficiency of PCR.However,it is ignored that DMSO can also decrease the amplification of specific sequences and result in non- specific amphfications.Non-specific bands are detected at 6% DMSO,as specific amplification declines.This is the first report about the effects of DMSO on the specificity of PCR and establishment of the method for avoiding non-specific products,which can be overcome by increasing template-primer ratio rather than annealing temperature.Our data will be of much help for random amphfied polymorphic DNA (RAPD) assays for molecular genetic analyses.