一种简便有效的PCR扩增片段直接回收克隆方法

A simple and efficient method for cloning PCR products

作者提供一种简便快速的克隆方法,即直接对琼脂糖凝胶电泳分离的PCR产物进行克隆,可获得较高的阳性克隆率.结果显示,300～700 bp大小的片段阳性克隆率为70.72%,300 bp以下和大于700 bp的片段阳性克隆率分别达到60.38%和45.33%.该方法适用于含有大量不同大小DNA片段的PCR产物并需要同时对其进行回收和克隆的样品,不仅可以大大简化实验过程,也可以降低假阳性出现的机会.

A simple method was introduced here for cloning a lot of PCR fragments with T vector in one time. A high positive rate of recombinant plasmid was gained. The positive rate of cloning for DNA fragments with 300-700 bp was 70.72%, while the rates for those shorter than 300 bp and longer than 700 bp were 60.38 % and 45.33 %, respectively.