通过双重聚合酶链反应进行的定点诱变
Site-directed mutagenesis by double polymerase chain reaction.
作者信息
Barik S
机构信息
Department of Biochemistry and Molecular Biology, College of Medicine, University of South Alabama, Mobile 36688-0002, USA.
出版信息
Mol Biotechnol. 1995 Feb;3(1):1-7. doi: 10.1007/BF02821329.
A simple and versatile method of in vitro site-specific mutagenesis based on polymerase chain reaction (PCR) is described. The complete method required the use of three oligonucleotide primers and two PCRs. The product of the first PCR was used as one of the primers (megaprimer) in the second PCR. Essentially 100% of the final product incorporated the desired mutation. The various aspects of the procedure and its application is described in detail.
描述了一种基于聚合酶链反应（PCR）的简单且通用的体外位点特异性诱变方法。完整的方法需要使用三种寡核苷酸引物和两次PCR。第一次PCR的产物用作第二次PCR中的一种引物（大引物）。基本上100%的最终产物都引入了所需的突变。详细描述了该程序的各个方面及其应用。
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