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Chemical synthesis of the mouse mitochondrial genome

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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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

Authors

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 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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