利用梯度PCR和降落PCR扩增CIRP基因的比较

摘要

提取SD大鼠总RNA,同时应用降落PCR和梯度PCR在常规PCR反应体系和低模板浓度PCR反应体系中扩增CIRP基因片段,对两种方法的扩增效果进行比较。结果表明：降落PCR法可以通过一次反应即顺利扩增出CIRP基因特异性条带,并且在模板浓度降低后也能扩增出产物;梯度PCR法扩增结果不稳定,不但有非特异性条带出现而且不能检测出低浓度模板的存在。结论：降落PCR省略了梯度PCR摸索最适退火温度的过程,并且其灵敏度和特异性均高于梯度PCR。%To extract the total RNA from rats,then to target CIRP genes by touch down PCR（TD-PCR） and gradient PCR（G-PCR） with the popular reaction system and the reaction system content low primer.The results indicated that TD-PCR was efficient in amplifying the CIRP gene from rats and it was more specific than G-PCR.TD-PCR with the same reaction system has higher specificity than G-PCR.It can efficiently improve specificity of PCR and be used to clone gene fragments which are difficult to clone by the G-PCR method.