噬菌体cDNA文库筛选方法的改良  

Improvement on phage cDNA library screening

作　　者：尚毅[1] 马璐琳[1] 亓增军[1] 刘大钧[1] 王秀娥[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,南京210095

出　　处：《生物学杂志》2010年第5期91-93,共3页Journal of Biology

基　　金：国家自然科学基金重点项目(编号30330380);美国McKnight基金CCRP项目资助

摘　　要：cDNA文库的构建和简便、快速的筛选是获得全长基因的重要途径,基于PCR的筛库方法具有快捷、灵敏的特点。研究改进了基于PCR的噬菌体cDNA文库筛选方法,用液体分装的方法,替代了文库筛选的关键步骤——涂板分区,省去了噬菌体文库铺平板、浸染、培养、划块洗脱的操作过程,使筛库的工作量减少,进一步提高了筛选速度和获得阳性克隆的效率。The construction of a cDNA library and establishment of a quick,simple and high-effective screening method is critical for obtaining the full length genes.PCR based cDNA library screening method is regarded to be fast and sensitive.In the present paper,we improved the PCR-based procedure of screening phage cDNA library.The key step in screening library,＂plate-division＂ was replaced by ＂partition in liquid＂,which avoids tedious plating,infection and washing procedures.This modification not only decreased the workload of library screening,but also improved the speed and efficiency of obtaining the positive clones.

关 键 词：噬菌体cDNA文库 PCR 文库筛选 液体分装 

分 类 号：Q939.48[生物学—微生物学]