Recombination-mediated PCR-directed plasmid construction in vivo in yeast

Kevin R. Oldenburg; Kham T. Vo; Susan Michaelis; Chris Paddon

doi:10.1093/nar/25.2.451

Recombination-mediated PCR-directed plasmid construction in vivo in yeast

Kevin R. Oldenburg, Kham T. Vo, Susan Michaelis, Chris Paddon

School of Medicine

Research output: Contribution to journal › Article › peer-review

381 Scopus citations

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.

Original language	English (US)
Pages (from-to)	451-452
Number of pages	2
Journal	Nucleic acids research
Volume	25
Issue number	2
DOIs	https://doi.org/10.1093/nar/25.2.451
State	Published - 1997

ASJC Scopus subject areas

Genetics

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title = "Recombination-mediated PCR-directed plasmid construction in vivo in yeast",

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.",

author = "Oldenburg, {Kevin R.} and Vo, {Kham T.} and Susan Michaelis and Chris Paddon",

note = "Funding Information: We would like to thank Walter Schmidt and Peter Schatz for comments on the manuscript. S.M. is funded by grants GM51508 and GM41223 from the National Institutes of Health.",

year = "1997",

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AU - Oldenburg, Kevin R.

AU - Vo, Kham T.

AU - Michaelis, Susan

AU - Paddon, Chris

N1 - Funding Information: We would like to thank Walter Schmidt and Peter Schatz for comments on the manuscript. S.M. is funded by grants GM51508 and GM41223 from the National Institutes of Health.

PY - 1997

Y1 - 1997

N2 - 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.

AB - 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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JF - Nucleic Acids Research

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Recombination-mediated PCR-directed plasmid construction in vivo in yeast

Abstract

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