Abstract
We describe a one-step, isothermal assembly method for synthesizing DNA molecules from overlapping oligonucleotides. The method cycles between in vitro recombination and amplification until the desired length is reached. As a demonstration of its simplicity and robustness, we synthesized the entire 16.3-kilobase mouse mitochondrial genome from 600 overlapping 60-mers.
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Acknowledgements
We thank Synthetic Genomics, Inc. for funding this work, J. Glass, Y.-H. Rogers, J. Gill, M. Frazier, E. Eisenstadt and M. Gibson for helpful discussions, and M. Algire and J. Zaveri for technical assistance.
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D.G.G. and C.M. designed research, performed research, analyzed data and wrote the paper. H.O.S., C.A.H. III and J.C.V. designed research and analyzed data.
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J.C.V. is chief executive officer and co-chief scientific officer of Synthetic Genomics, Inc (SGI). H.O.S. is co-chief scientific officer and a member of the board of directors of SGI. C.A.H. III is chairman of the SGI Scientific Advisory Board. J.C.V., H.O.S. and C.A.H. III hold SGI stock.
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Gibson, D., Smith, H., Hutchison, C. et al. Chemical synthesis of the mouse mitochondrial genome. Nat Methods 7, 901–903 (2010). https://doi.org/10.1038/nmeth.1515
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DOI: https://doi.org/10.1038/nmeth.1515
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