A method for multi-site-directed mutagenesis based on homologous recombination
A method for multi-site-directed mutagenesis based on homologous recombination
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DOI：
10.1016/j.ab.2012.05.002
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发表时间：
2012-08-01
影响因子：
2.9
通讯作者：
Katzen, Federico
中科院分区：
文献类型：
--
作者：
Liang, Xiquan;Peng, Lansha;Katzen, Federico
我们开发了一种有效的方法，通过同源重组同时在一个质粒DNA中引入多达三个突变。该策略与各种突变兼容，包括不同大小的质粒中的简并密码子。与其他方法相比，该方法使用相同的试剂集进行单点和多点突变分析，最大限度地减少了所需的程序步骤，并显示出非常高的诱变效率。(C)2012 Elsevier Inc.保留所有权利。
We have developed an efficient method for the simultaneous introduction of up to three mutations in a plasmid DNA via homologous recombination. The strategy is compatible with a variety of mutations, including degenerate codons in plasmids of different sizes. In contrast to other methodologies, this approach employs the same set of reagents for both single- and multi-site mutagenesis assays, minimizes the required protocol steps, and exhibits remarkably high mutagenesis efficiencies. (C) 2012 Elsevier Inc. All rights reserved.