A method for multi-site-directed mutagenesis based on homologous recombination

作者：Liang Xiquan; Peng Lansha; Li Ke; Peterson Todd; Katzen Federico^*

来源：Analytical Biochemistry, 2012, 427(1): 99-101.

DOI：10.1016/j.ab.2012.05.002

• Mutagenize
• Oligonucleotide
• Plasmid
• Mutagenic
• Primer
• PCR

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.

• 出版日期2012-8-1