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Science. 2005 Sep 9;309(5741):1728-32. Epub 2005 Aug 4.

Accurate multiplex polony sequencing of an evolved bacterial genome.

Author information

1
Department of Genetics, Harvard Medical School, Boston, MA 02115, USA. shendure@alumni.princeton.edu

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.

PMID:
16081699
DOI:
10.1126/science.1117389
[Indexed for MEDLINE]
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