A method for multi-site-directed mutagenesis based on homologous recombination
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- 作者：Xiquan Liang ; Lansha Peng ; Ke Li ; Todd Peterson ; Federico Katzen
- 关键词：Mutagenize ; Oligonucleotide ; Plasmid ; Mutagenic ; Primer ; PCR
- 刊名：Analytical Biochemistry
- 出版年：2012
- 出版时间：1 August, 2012
- 年：2012
- 卷：427
- 期：1
- 页码：99-101
- 全文大小：289 K
文摘
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.