一种快速的重组农杆菌PCR鉴定方法  

A Rapid Identification Method of Recombinant Agrobacterium PCR Reaction

作　　者：周晓馥[1] 赵晶[1] 赵航[1] 何珊珊[1] 高旭伟[1] 张冰[1] 徐洪伟[1] 

机构地区：[1]吉林师范大学植物资源科学与绿色生产吉林省重点实验室,吉林四平136000

出　　处：《吉林师范大学学报（自然科学版）》2015年第2期109-111,共3页Journal of Jilin Normal University:Natural Science Edition

基　　金：国家自然科学基金项目(31070224);吉林省科技发展计划项目(20140101136JC)

摘　　要：建立一种快速的重组农杆菌PCR扩增鉴定方法,采用煮沸法处理重组农杆菌,使农杆菌高温裂解,释放质粒DNA,直接进行PCR扩增反应,检测目的基因片段.结果表明,重组质粒PCR扩增效果较好,目标条带较为明显.通过煮沸重组农杆菌,进行PCR扩增反应鉴定质粒DNA,不仅实验操作简单安全,缩短实验时间,并且避免了酚、氯仿试剂在提取质粒DNA过程中的残留对后续分子鉴定质量的影响.Boiling method was introduced to establish a rapid Recombinant Agrobacterium identification method. Recombinant Agrobacterium was boiled to release the plasmid DNA. After that,PCR reaction was performed,in order to amplify specific DNA sequences. Our results indicated that the purity of the amplified product was good and the target stripe was obvious. It was easy,safe and time-saving to identify plasmid DNA via boiling and PCR method. At the same time,it avoided the phenol and chloroform residue from subsequent molecular identification.

关 键 词：重组发根农杆菌 煮沸法 PCR扩增反应 

分 类 号：Q943.1[生物学—植物学]