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Nucleic Acids Research Advance Access published online on September 10, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp687
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Synthetic Biology and Chemistry

Synthesis of DNA fragments in yeast by one-step assembly of overlapping oligonucleotides

Daniel G. Gibson*

The J. Craig Venter Institute, Synthetic Biology Group, 9704 Medical Center Drive, Rockville, MD 20850, USA

*To whom correspondence should be addressed. Tel: +1 301 795 7268; Fax: +1 240 268 4004; Email: dgibson{at}jcvi.org

Received January 1, 2009. Revised August 1, 2009. Accepted August 4, 2009.

Here it is demonstrated that 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. This straightforward scheme for assembling chemically-synthesized oligonucleotides could be a useful tool for building synthetic DNA molecules.


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