Molecular biology: Accurate multiplex polony sequencing of an evolved bacterial genome

Jay Shendure, Gregory J. Porreca, Nikos B. Reppas, Xiaoxia Lin, John P. McCutcheon, Abraham M. Rosenbaum, Michael D. Wang, Kun Zhang, Robi D. Mitra, George M. Church

Research output: Contribution to journalArticle

932 Scopus citations

Abstract

We describe a DNA sequencing technology in which a commonly available, inexpensive epifluorescence microscope is converted to rapid nonelectrophoretic DNA sequencing automation. We apply this technology to resequence an evolved strain of Escherichia coli at less than one error per million consensus bases. A cell-free, mate-paired library provided single DNA molecules that were amplified in parallel to 1-micrometer beads by emulsion polymerase chain reaction. Millions of beads were immobilized in a polyacrylamide gel and subjected to automated cycles of sequencing by ligation and four-color imaging. Cost per base was roughly one-ninth as much as that of conventional sequencing. Our protocols were implemented with off-the-shelf instrumentation and reagents.

Original languageEnglish
Pages (from-to)1728-1732
Number of pages5
JournalScience
Volume309
Issue number5741
DOIs
StatePublished - Sep 9 2005

Fingerprint Dive into the research topics of 'Molecular biology: Accurate multiplex polony sequencing of an evolved bacterial genome'. Together they form a unique fingerprint.

  • Cite this

    Shendure, J., Porreca, G. J., Reppas, N. B., Lin, X., McCutcheon, J. P., Rosenbaum, A. M., Wang, M. D., Zhang, K., Mitra, R. D., & Church, G. M. (2005). Molecular biology: Accurate multiplex polony sequencing of an evolved bacterial genome. Science, 309(5741), 1728-1732. https://doi.org/10.1126/science.1117389