Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome

• DOI: 10.1126/science.1117389
• Publication Date: 2005-08-05T00:44:05Z
• Authors: 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
• Keywords: Multiplex

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