小提质粒快速排除假阳性克隆的新方法

Rapid Mini-Preparation of Plasmid DNA for Eliminating False Positive Clones

对传统的小提质粒方法进行了简化,在15 min内提出重组子质粒,从而快速地排除不含质粒的假阳性菌落。为验证方法的可靠性,构建了7个重组子,并应用PCR方法进一步对筛选结果进行鉴定。结果表明:此方法简便、迅速、可靠、重复性好,适用于革兰氏阴性菌,可以先排除一部分假阳性克隆,缩小筛选范围。

A rapid method was developed to greatly simplify the procedures of plasmid DNA purification within about 15 min and greatly reduce the experiment cost compared with traditional method. Seven recombinants were constructed and identified by PCR in order to confirm reliability of the new method.The results indicated that this new method was simple and inexpensive and suitable for gram-negative bacterium.It can be used for rapidly eliminating false positive clones.