SITE-DIRECTED MUTAGENESIS BY DOUBLE POLYMERASE CHAIN-REACTION
SITE-DIRECTED MUTAGENESIS BY DOUBLE POLYMERASE CHAIN-REACTION
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DOI：
10.1007/bf02821329
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发表时间：
1995-02-01
影响因子：
2.6
通讯作者：
BARIK, S
中科院分区：
文献类型：
--
作者：
BARIK, S
本文介绍了一种基于聚合酶链反应（PCR）的体外定点突变方法。完整的方法需要使用三个寡核苷酸引物和两个PCR。第一次PCR的产物用作第二次PCR中的引物之一（大引物）。基本上100%的最终产物掺入了所需的突变。详细描述了该程序的各个方面及其应用。
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.