直接用细菌重组体进行聚合酶链反应及其在克隆鉴定中的应用

作用不需提纯ＤＮＡ模，可直接从质粒的宿主菌中扩增插入片段的方法进行扩增和克隆鉴定。结果说明，该方法不但在扩增的灵敏度和特异性与经用酚氯仿抽提纯化的质粒ＤＮＡ作模板一致，使质粒鉴定的时间由原来４８－７２ｈ缩短至２－８ｈ，而且节省了细胞培养及质粒纯化过所用的试剂及器械。特别是对于无法用酶切法鉴定的克隆有特殊意义。

Direct polymerase chain reaction using recombinant bacteria as template and its application in identification of recombinant clones

The present paper reports a new method for identification of recombmant plasmid. The protocol involved using overnight cultured bacteria or transforming colony as template directly and purified plasmid DNA as controls. The results showed that the amplified products of bacteria were almost identical to those of purified plasmid DNA. It was found that this procedure curtailed the time identification of recombinant clones from 2~3 d to 2~8 h, for while the cost was much reduced. The findings demonstrated that the present method of identifying recombinant clones is very useful for clones without recognization sites.