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Chemical synthesis of the mouse mitochondrial genome

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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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Figure 1: Schematic demonstrating assembly of the synthetic mouse mitochondrial genome.
Figure 2: Summary of results for obtaining the 75 sequence-verified first-stage assemblies.

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GenBank/EMBL/DDBJ

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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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Authors and Affiliations

  1. The J. Craig Venter Institute, Rockville, Maryland, USA

    Daniel G Gibson, J Craig Venter & Chuck Merryman

  2. The J. Craig Venter Institute, San Diego, California, USA

    Hamilton O Smith, Clyde A Hutchison III & J Craig Venter

Authors
  1. Daniel G Gibson
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  2. Hamilton O Smith
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  3. Clyde A Hutchison III
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  4. J Craig Venter
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  5. Chuck Merryman
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Contributions

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.

Corresponding authors

Correspondence to Daniel G Gibson or Chuck Merryman.

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Competing interests

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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Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Tables 1–15, Supplementary Note 1 (PDF 1416 kb)

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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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