一种PCR快速鉴定重组体DNA阳性克隆及插入方向的方法  

Rapid Determination of Foreign DNA Orientation in Qoned Vector

作　　者：廖世奇[1] 张春梅[1] 张雪力[1] 孙雨葳 王黎[1] 王晓辉[1] 

机构地区：[1]甘肃省医学科学研究院,兰州730050 [2]青海省妇产儿童医院,西宁810007

出　　处：《科学技术与工程》2004年第2期89-90,96,共3页Science Technology and Engineering

基　　金：国家自然科学基金(30271219)

摘　　要：当外源DNA以非定向的方式插入克隆载体时,需用测序或酶切的方法确定插入方向,上述两种方法或昂贵或费力。描述了一种快速、简便的PCR法,即利用载体的通用引物和目的片段的两个特异性引物直接扩增重组菌落,从而达到鉴别DNA插入方向的目的。Generally, DNA sequencing or asymmetrical digest of recombinant plasmids must be done to determine the orientation of foreign DNA fragments in cloned vector. The former is expensive and the later proves difficult in some cases. A rapid and simple PCR-based method to solve the problem is described. The orientation of the insert DNA is defined by direct PCR amplification of bacterial colonies using a universal primer complemented to the T7 pro-motor of the vector and two insert specific primers.

关 键 词：外源DNA 插入方向 载体通用引物 PCR快速鉴定 克隆 分子生物学 

分 类 号：Q785[生物学—分子生物学]