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Nature. 2005 Sep 15;437(7057):376-80. Epub 2005 Jul 31.

Genome sequencing in microfabricated high-density picolitre reactors.

Author information

  • 1454 Life Sciences Corp., 20 Commercial Street, Branford, Connecticut 06405, USA.

Erratum in

  • Nature. 2006 May 4;441(7089):120. Ho, Chun He [corrected to Ho, Chun Heen].

Abstract

The proliferation of large-scale DNA-sequencing projects in recent years has driven a search for alternative methods to reduce time and cost. Here we describe a scalable, highly parallel sequencing system with raw throughput significantly greater than that of state-of-the-art capillary electrophoresis instruments. The apparatus uses a novel fibre-optic slide of individual wells and is able to sequence 25 million bases, at 99% or better accuracy, in one four-hour run. To achieve an approximately 100-fold increase in throughput over current Sanger sequencing technology, we have developed an emulsion method for DNA amplification and an instrument for sequencing by synthesis using a pyrosequencing protocol optimized for solid support and picolitre-scale volumes. Here we show the utility, throughput, accuracy and robustness of this system by shotgun sequencing and de novo assembly of the Mycoplasma genitalium genome with 96% coverage at 99.96% accuracy in one run of the machine.

Comment in

PMID:
16056220
[PubMed - indexed for MEDLINE]
PMCID:
PMC1464427
Free PMC Article

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