一种用于双链质粒DNA定点诱变的简单高效方法。
A simple and efficient method for site-directed mutagenesis with double-stranded plasmid DNA.
作者信息
Lai D, Zhu X, Pestka S
机构信息
Department of Molecular Genetics and Microbiology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854-5635.
出版信息
Nucleic Acids Res. 1993 Aug 25;21(17):3977-80. doi: 10.1093/nar/21.17.3977.
Abstract
A general, simple and efficient method for preparing site-specific mutations in double-stranded plasmid DNA without the need for special plasmids, bacterial strains or reagents is described. Only one synthetic oligonucleotide for each mutation is required, subcloning is unnecessary and a high efficiency of mutation (58-97%) was obtained. If two synthetic oligonucleotide primers are used, two separate mutations can be simultaneously created in a single reaction tube.
摘要
本文描述了一种通用、简单且高效的方法，用于在双链质粒DNA中制备位点特异性突变，无需特殊质粒、细菌菌株或试剂。每个突变仅需一个合成寡核苷酸，无需亚克隆，且获得了较高的突变效率（58-97%）。如果使用两个合成寡核苷酸引物，则可在单个反应管中同时产生两个独立的突变。
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