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

@article{Liang2012AMF,
  title={A method for multi-site-directed mutagenesis based on homologous recombination.},
  author={Xiquan Liang and Lansha Peng and Ke Li and Todd C. Peterson and Federico Katzen},
  journal={Analytical biochemistry},
  year={2012},
  volume={427 1},
  pages={99-101}
}
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. 
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