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Science. 2008 Apr 4;320(5872):106-9. doi: 10.1126/science.1150427.

Single-molecule DNA sequencing of a viral genome.

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  • 1Helicos BioSciences Corporation, One Kendall Square, Cambridge, MA 02139, USA. tharris@helicosbio.com

Abstract

The full promise of human genomics will be realized only when the genomes of thousands of individuals can be sequenced for comparative analysis. A reference sequence enables the use of short read length. We report an amplification-free method for determining the nucleotide sequence of more than 280,000 individual DNA molecules simultaneously. A DNA polymerase adds labeled nucleotides to surface-immobilized primer-template duplexes in stepwise fashion, and the asynchronous growth of individual DNA molecules was monitored by fluorescence imaging. Read lengths of >25 bases and equivalent phred software program quality scores approaching 30 were achieved. We used this method to sequence the M13 virus to an average depth of >150x and with 100% coverage; thus, we resequenced the M13 genome with high-sensitivity mutation detection. This demonstrates a strategy for high-throughput low-cost resequencing.

PMID:
18388294
[PubMed - indexed for MEDLINE]
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