农杆菌质粒DNA提取方法的优化  

A Modified Method for Isolation of Plasmid DNA from Agribaterium tumefacien

作　　者：王友如[1] 崔洪志[2] 郭三堆[2] 

机构地区：[1]湖北师范学院生物系,湖北黄石435002 [2]中国农业科学院生物技术所基因工程实验室,北京100081

出　　处：《生物技术》2005年第4期41-43,共3页Biotechnology

摘　　要：目的:为了减化操作过程,减少药品对操作人员有直接或间接危害,提高农杆菌质粒DNA的产率和实验结果的稳定性。方法:对常规碱裂解法进行了改动,改进的方法通过加大菌液的收集量,利用LiCl沉淀RNA,简化操作流程和严格反映环境条件,并且在操作过程中去除了酚、氯仿等对人体有害的试剂。结果:提取时间缩短,结果稳定,产率在1.5μgμL以上。结论:用这种方法提取的质粒可以满足大多数分子生物学常规实验如DNA的酶切、PCR鉴定的要求。Objective： To simplify the operating process reduce the direct or indirect harm by medicine to reseachers , and finally improve plasmid DNA productivity and the stablity of the result. Method： Modify the alkaline lysis method for isolation of plasmid DNA from Agribateriium tumefacien, increase the volume of gathered Agribaterium tumefacie, and sediment RNA by LiCl, siniplify the operating process , anti remove phenol and chloroform which is harmful io reseachers. Result： the extrcted time has been shortened, the results are stable , and the plasmid DNA isolated by the modified method often contamined with large amount of RNA at the level of 1.5μg/μL . Conclusion：The modified method for isolation of plasmid DNA from Agribaterium tumefacien,which is no harmful to human beings, is more suitable to test recombinant plasmid DNA with restrietion endonuclease and PCR testing method.

关 键 词：农杆菌 质粒DNA 提取方法优化 

分 类 号：Q782[生物学—分子生物学]