Reference: Alcalde M (2010) Mutagenesis protocols in Saccharomyces cerevisiae by in vivo overlap extension. Methods Mol Biol 634:3-14

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Abstract


A high recombination frequency and its ease of manipulation has made Saccharomyces cerevisiae a unique model eukaryotic organism to study homologous recombination. Indeed, the well-developed recombination machinery in S. cerevisiae facilitates the construction of mutant libraries for directed evolution experiments. In this context, in vivo overlap extension (IVOE) is a particularly attractive protocol that takes advantage of the eukaryotic apparatus to carry out combinatorial saturation mutagenesis, site-directed recombination or site-directed mutagenesis, avoiding ligation steps and additional PCR reactions that are common to standard in vitro protocols.

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Journal Article | Research Support, Non-U.S. Gov't
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Alcalde M
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