【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。更多还原

【Abstract】 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.更多还原