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ReportsSingle-Molecule DNA Sequencing of a Viral GenomeThe 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.
1 Helicos BioSciences Corporation, One Kendall Square, Cambridge, MA 02139, USA. * To whom correspondence should be addressed. E-mail: tharris{at}helicosbio.com
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Science. ISSN 0036-8075 (print), 1095-9203 (online)