兼并PCR技术反应条件的探索  

Studies on the PCR conditions of Degenerate Primer

作　　者：吴茜 敬永升[2] 王艺霖 刘肖肖 王洪涛[1] WU Xi;JING Yongsheng;WANG Yilin;LIU Xiaoxiao;WANG Hongtao(College of Life Science,Henan University,Kaifeng 475004,China;College of pharmacy,Henan University,Kai feng 475004,China;College of Life Science,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]河南大学生命科学学院,河南开封475004 [2]河南大学药学院,河南开封475004 [3]南京农业大学生命科学学院,南京210095

出　　处：《河南大学学报（医学版）》2017年第4期272-276,共5页Journal of Henan University:Medical Science

基　　金：中国烟草总公司标准预研项目(2013QB017)

摘　　要：[目的]探索兼并PCR技术反应的条件。[方法]依据过氧化物酶N-末端氨基酸序列设计的兼并引物,提取幼龄梨果实的DNA和RNA并将RNA反转录为cDNA,分别以DNA、cDNA和PCR产物为模板,利用PCR和琼脂糖电泳分析的方法,在不同的退火温度和引物添加量的配比下,对PCR条件进行优化。[结果]优化了PCR反应的条件,并且得到目的基因。[结论]优化后的兼并PCR技术可以更好地应用于新基因的克隆、病毒的检测等研究领域。〔Objective〕Studies on the PCR conditions of degenerate Primer.〔Methods〕The PCR conditions were optimized by PCR and agarose gel electrophoresis at different annealing temperatures and primer addition,by designing the degenerate primers on the according of N terminal amino acid sequence of POD,and extracting the DNA and RNA from Pear fruitlets,and RNA was transcribed into cDNA.〔Results〕The conditions of PCR reaction were optimized,and the target gene was obtained.〔Conclusion〕The optimized PCR of degenerate Primer technology can be applied to the cloning of new genes,detection of viruses and other research fields.

关 键 词：兼并引物 PCR 过氧化物酶 退火温度 引物添加量的配比 

分 类 号：Q78[生物学—分子生物学]