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

Science (Washington, DC, United States) (2005), 309(5741), 1728-1732 CODEN: SCIEAS; ISSN: 0036-8075. English.

The authors describe a DNA sequencing technol. in which a commonly available, inexpensive epifluorescence microscope is converted to rapid nonelectrophoretic DNA sequencing automation. The authors apply this technol. 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 mols. that were amplified in parallel to 1-μm 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. The protocols were implemented with off-the-shelf instrumentation and reagents.