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Recombination-mediated PCR-directed plasmid construction in vivo in yeast
K R Oldenburg1, K T Vo, S Michaelis
1Affymax Research Institute, 4001 Miranda Avenue, Palo Alto, CA 94304, USA.
Nucleic Acids Research
|January 15, 1997
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.