利用梯度PCR和降落PCR扩增CIRP基因的比较  

Comparison of the Amplification of CIRP Genes with Touch Down PCR and Gradient PCR

作　　者：李玉恒[1] 赵明慧[1] 赵紫阳[1] 李峥[1] 袁雪[1] 李昌盛[1] 林增[1] 胡婧[1] 李俭[1] 李士泽[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319

出　　处：《黑龙江八一农垦大学学报》2011年第5期46-49,75,共5页journal of heilongjiang bayi agricultural university

基　　金：农业部948重点项目(2011-G35);2011年黑龙江省研究生创新科研项目(YJSCX2011-267HLJ)

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.

关 键 词：降落PCR 梯度PCR 退火温度 CIRP基因 

分 类 号：Q78[生物学—分子生物学]