浙江医学2017，Vol.39Issue(14)：1166-1169,后插2,5.DOI:10.12056/j.issn.1006-2785.2017.39.14.2017-1035
利用退火反应产生粘性末端的基因克隆新方法
A new method for gene clone producing cohesive terminals by annealingreaction
摘要
Abstract
Objective To develop a new effective gene clone method by designing three or four primers to produce cohesive terminals.Methods Two independent PCRs were run based on these designed primers,mixed the two PCR products together,and then went through denaturation and annealing.After that,the mixed products were ligated with the digested vector and the ligation products were transformed into DH5 α competent cells.Results The target genes (P90rsk,Nprl2,Mios,Ragc,S6k,Pkca) were constructed on vector pcDNA3.1 (+),which were digested with EcoRV and Notl.The constructedgenes were verified by single restriction digestion and sequencing.Conclusion The developed method overcomes the limitation of restriction endonuclease sites present in the target genes and can also omit the digestion process of the PCR products and subsequent purification.关键词
分子克隆/聚合酶链反应/退火/限制性酶切位点/粘性末端Key words
Molecular cloning/Polymerase chain reaction/Annealing/Restriction site/Cohesive end引用本文复制引用
赵婷婷,曾静静,王启军,褚茂平..利用退火反应产生粘性末端的基因克隆新方法[J].浙江医学,2017,39(14):1166-1169,后插2,5.基金项目
浙江省自然科学基金资助项目(LZ17H250001) （LZ17H250001）
