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Nucleic Acids Research 2004 32(20):e162; doi:10.1093/nar/gnh160
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Published online 23 November 2004

Nucleic Acids Research, Vol. 32 No. 20 © Oxford University Press 2004; all rights reserved

Protein-mediated error correction for de novo DNA synthesis

Peter A. Carr1,2, Jason S. Park3, Yoon-Jae Lee4, Tiffany Yu5, Shuguang Zhang6 and Joseph M. Jacobson1,2,*

1 Center for Bits and Atoms, 2 Media Laboratory, 3 Department of Mechanical Engineering, 4 Department of Biology, 5 Department of Chemical Engineering and 6 Center for Biomedical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA

* To whom correspondence should be addressed. Tel: +1 617 253 7209; Fax: +1 617 258 6264; Email: jacobson{at}media.mit.edu

Received October 8, 2004; Revised and Accepted October 30, 2004

The availability of inexpensive, on demand synthetic DNA has enabled numerous powerful applications in biotechnology, in turn driving considerable present interest in the de novo synthesis of increasingly longer DNA constructs. The synthesis of DNA from oligonucleotides into products even as large as small viral genomes has been accomplished. Despite such achievements, the costs and time required to generate such long constructs has, to date, precluded gene-length (and longer) DNA synthesis from being an everyday research tool in the same manner as PCR and DNA sequencing. A critical barrier to low-cost, high-throughput de novo DNA synthesis is the frequency at which errors pervade the final product. Here, we employ a DNA mismatch-binding protein, MutS (from Thermus aquaticus) to remove failure products from synthetic genes. This method reduced errors by >15-fold relative to conventional gene synthesis techniques, yielding DNA with one error per 10 000 base pairs. The approach is general, scalable and can be iterated multiple times for greater fidelity. Reductions in both costs and time required are demonstrated for the synthesis of a 2.5 kb gene.


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