An improved PCR-mutagenesis strategy for two-site mutagenesis or sequence swapping between related genes
An improved PCR-mutagenesis strategy for two-site mutagenesis or sequence swapping between related genes
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DOI：
10.1093/nar/26.7.1848
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发表时间：
1998-04-01
影响因子：
14.9
通讯作者：
Joly, E
中科院分区：
文献类型：
--
作者：
Kirsch, RD;Joly, E
QuikChange(TM) 方案是最简单、最快的定点诱变方法之一，但一次仅在一个位点引入突变，并且需要两种 HPLC 纯化的互补寡核苷酸。在此，我们描述该方法可与非重叠寡核苷酸一起使用。通过这样做，可以同时诱变两个单独的位点，或者可以通过使用 第二个“标准”寡核苷酸，通过进一步修改，我们还使用 QuikChange(TM) 方法在密切相关的基因之间交换 DNA 序列。
The QuikChange(TM) protocol is one of the simplest and fastest methods for site-directed mutagenesis, but introduces mutations at only one site at a time, and requires two HPLC-purified complementary oligonucleotides, Here, we describe that this method can be used with non-overlapping oligonucleotides, By doing this, two separate sites can be mutagenised simultaneously, or money can be saved by using a second 'standard' oligonucleotide, By a further modification, we have also used the QuikChange(TM) approach to exchange DNA sequences between closely related genes.