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Abstract

We synthesized reversible terminators with tethered inhibitors for next-generation sequencing. These were efficiently incorporated with high fidelity while preventing incorporation of additional nucleotides, and we used them to sequence canine bacterial artificial chromosomes in a single-molecule system that provided even coverage for over 99% of the region sequenced. This single-molecule approach generated high-quality sequence data without the need for target amplification and thus avoided concomitant biases.

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Acknowledgements

We thank all of the past and present colleagues at Helicos who have contributed to this work. This work was made possible by grant number R01 HG004144 from the National Human Genetics Research Institute (NHGRI) at the US National Institutes of Health. Contents of this paper are solely the responsibility of the authors and do not necessarily represent the official views of NHGRI.

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Affiliations

  1. Helicos BioSciences Corporation, Cambridge, Massachusetts, USA.

    • Jayson Bowers
    • , Judith Mitchell
    • , Eric Beer
    • , Philip R Buzby
    • , Marie Causey
    • , J William Efcavitch
    • , Mirna Jarosz
    • , Edyta Krzymanska-Olejnik
    • , Li Kung
    • , Doron Lipson
    • , Geoffrey M Lowman
    • , Subramanian Marappan
    • , Peter McInerney
    • , Adam Platt
    • , Atanu Roy
    • , Suhaib M Siddiqi
    • , Kathleen Steinmann
    •  & John F Thompson

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Contributions

J.M., J.W.E., E.K.-O., S.M., A.R. and S.M.S. synthesized and/or designed new compounds. J.B., E.B. and P.M. designed and executed biochemical assays. M.C., M.J., L.K., G.M.L., A.P. and K.S. sequenced DNA samples. M.C., M.J., L.K., D.L., G.M.L. and A.P. analyzed sequencing data. J.B., J.M., E.K.-O., S.M.S. and J.F.T. wrote the manuscript. J.B., P.R.B., M.C., J.W.E., M.J. and S.M.S. supervised and managed the project.

Competing interests

J.B, J.M., E.B., P.R.B., M.C., J.W.E., M.J., E.K.-O., L.K., D.L., G.M.L., S.M., P.M., A.P., A.R., S.M.S., K.S. and J.F.T. are present or former employees of Helicos BioSciences.

Corresponding author

Correspondence to John F Thompson.

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DOI

https://doi.org/10.1038/nmeth.1354

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