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, 25 (2), 451-2

Recombination-mediated PCR-directed Plasmid Construction in Vivo in Yeast

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Recombination-mediated PCR-directed Plasmid Construction in Vivo in Yeast

K R Oldenburg et al. Nucleic Acids Res.

Abstract

We have extended the technique of PCR-directed recombination in Saccharomyces cerevisiae to develop a simple method for plasmid or gene construction in the absence of suitable restriction sites. The DNA to be cloned is PCR-amplified with 30-40 bp of homology to a linearized yeast plasmid. Co-transformation into yeast results in homologous recombination at a position directed by the PCR oligonucleotides.

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