Simple, robust methods for high-throughput nanoliter-scale DNA sequencing

  1. Duane E. Smailus1,
  2. Andre Marziali2,
  3. Philip Dextras2,
  4. Marco A. Marra1, and
  5. Robert A. Holt1,3
  1. 1 Canada's Michael Smith Genome Sciences Centre, British Columbia Cancer Agency, Vancouver, British Columbia, Canada V5Z 4S6
  2. 2 Department of Physics and Astronomy, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z1.

Abstract

We have developed high-throughput DNA sequencing methods that generate high quality data from reactions as small as 400 nL, providing an approximate order of magnitude reduction in reagent use relative to standard protocols. Sequencing of clones from plasmid, fosmid, and BAC libraries yielded read lengths (PHRED20 bases) of 765 ± 172 (n = 10,272), 621 ± 201 (n = 1824), and 647 ± 189 (n = 568), respectively. Implementation of these procedures at high-throughput genome centers could have a substantial impact on the amount of data that can be generated per unit cost.

Footnotes

  • [Supplemental material is available online at www.genome.org.]

  • Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.4221805. Article published online before print in September 2005.

  • ↵3 Corresponding author. E-mail rholt{at}bcgsc.ca; fax (604) 877-6085.

    • Accepted August 1, 2005.
    • Received June 1, 2005.
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This Article

  1. Published in Advance September 16, 2005, doi: 10.1101/gr.4221805 Genome Res. 15: 1447-1450 Cold Spring Harbor Laboratory Press

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