Life Technologies, Carlsbad, CA 92008, USA.
Anal Biochem. 2012 Aug 1;427(1):99-101. doi: 10.1016/j.ab.2012.05.002. Epub 2012 May 10.
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.
我们开发了一种高效的方法，通过同源重组在质粒 DNA 中同时引入多达三个突变。该策略与各种突变兼容，包括不同大小质粒中的简并密码子。与其他方法相比，该方法在单一位点和多位点诱变实验中使用相同的试剂，最大限度地减少所需的实验步骤，并表现出极高的突变效率。
