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Published online before print September 16, 2005
Genome Research, DOI: 10.1101/gr.4221805
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Genome Research Vol 0, Issue 2005, gr.4221805, 2005
© 2005 Cold Spring Harbor Laboratory Press

Methods

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

Duane E. Smailus1, Andre Marziali2, Philip Dextras2, Marco A. Marra1 and Robert A. Holt1,3

1 Canada's Michael Smith Genome Sciences Centre, British Columbia Cancer Agency, Vancouver, British Columbia, Canada V5Z 4S6 , 2 Department of Physics and Astronomy, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z1

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.

[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.


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