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Methods Mol Biol. 2012;852:11-21. doi: 10.1007/978-1-61779-564-0_2.

Oligonucleotide assembly in yeast to produce synthetic DNA fragments.

Author information

  • 1Department of Synthetic Biology, J. Craig Venter Institute, Inc., Rockville, MD, USA. dgibson@jcvi.org

Abstract

The yeast Saccharomyces cerevisiae can take up and assemble at least 38 overlapping single-stranded oligonucleotides and a linear double-stranded vector in one transformation event. These oligonucleotides can overlap by as few as 20 bp and can be as long as 200 nucleotides in length to produce kilobase-sized synthetic DNA molecules. A protocol for designing the oligonucleotides to be assembled, transforming them into yeast, and confirming their assembly is described here. This straightforward scheme for assembling chemically synthesized oligonucleotides can be a useful tool for building synthetic DNA molecules.

PMID:
22328422
[PubMed - indexed for MEDLINE]
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