PCR在消减文库构建中的应用  

Applification of PCR in subtractive library of cDNA

作　　者：殷智榕[1] 高冬玲[1] 陈奎生[2] 姜国忠[1] 张云汉[1] 

机构地区：[1]河南医科大学第一附属医院病理科,郑州450052 [2]河南省肿瘤病理重点实验室,郑州450052

出　　处：《河南医科大学学报》2001年第2期165-166,共2页Journal of Henan Medical University

摘　　要：目的 :消减文库构建过程中 ,用PCR技术快速筛选重组阳性克隆。方法 :将白色单菌落加入氨苄抗性LB培养液中 ,37℃摇振培养过夜 ,取细菌悬液作PCR模板。结果和结论 :以PCR方法筛查重组阳性克隆 ,可以简便快速鉴定重组阳性克隆 ,不需提取质粒。筛选重组阳性克隆可直接用细菌悬液作PCR模板 ,在消减文库构建时 ,能大大提高工作效率。Aim: To develop a PCR technique for rapid screening of recombinant plasmid in subtractive library of cDNA.Methods:The recombinant colonies were transferred into LB culture medium with ampicillin,and shaken overnight at 37℃,then took the medium for PCR templet.Results and Conclusion :The PCR screening method was convenient and fast for confirming positive recombinant clone,and there was no need for preparation and purification of the plasmid.Recombinant clone can be analyzed directly with the use of PCR.It is very efficient,especially in establishment of subtractive library of cDNA

关 键 词：PCR 重组阳性克隆 消减文库 

分 类 号：Q785[生物学—分子生物学]