Saris P E, Paulin L
Biotechniques. 1990 Dec;9(6):694, 696-7.
We have developed a polymerase chain reaction (PCR)-based procedure to facilitate the selection of recombinant clones. The insert to be cloned is ligated to an antibiotic resistance marker. The ligation product is amplified by PCR, followed by standard cloning procedure into a bacterial vector. The selection for the antibiotic resistance coded by the PCR product ensures 100% insertion frequency, eliminating the screening of the transformants.
我们开发了一种基于聚合酶链反应(PCR)的方法,以促进重组克隆的筛选。待克隆的插入片段与抗生素抗性标记连接。连接产物通过PCR扩增,然后按照标准克隆程序克隆到细菌载体中。对PCR产物编码的抗生素抗性进行筛选可确保100%的插入频率,从而无需对转化体进行筛选。