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.
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Acknowledgments
The author would like to thank the Synthetic Biology Group at JCVI for the helpful discussions and Synthetic Genomics, Inc. for funding this work.
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Gibson, D.G. (2012). Oligonucleotide Assembly in Yeast to Produce Synthetic DNA Fragments. In: Peccoud, J. (eds) Gene Synthesis. Methods in Molecular Biology, vol 852. Humana Press. https://doi.org/10.1007/978-1-61779-564-0_2
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DOI: https://doi.org/10.1007/978-1-61779-564-0_2
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