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Gibson DG (2012) Oligonucleotide assembly in yeast to produce synthetic DNA fragments. Methods Mol Biol 852():11-21
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.
| Status: Published | Type: Journal Article | PubMed ID: 22328422 |
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