Single-molecule DNA sequencing of a viral genome

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


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Bacteriophage M13 / genetics*
  • Computational Biology / methods
  • DNA Primers
  • DNA, Viral / chemistry
  • DNA, Viral / genetics*
  • Genome, Viral*
  • Mutation
  • Sequence Alignment
  • Sequence Analysis, DNA / methods*
  • Software
  • Templates, Genetic


  • DNA Primers
  • DNA, Viral