Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome

@article{citeulike:320356, 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.}, author = {Shendure, Jay and Porreca, Gregory J. and Reppas, Nikos B. and Lin, Xiaoxia and Mccutcheon, John P. and Rosenbaum, Abraham M. and Wang, Michael D. and Zhang, Kun and Mitra, Robi D. and Church, George M. }, citeulike-article-id = {320356}, doi = {10.1126/science.1117389}, journal = {Science}, keywords = {genomics, sequencing}, month = {September}, number = {5741}, pages = {1728--1732}, posted-at = {2008-01-11 19:08:07}, priority = {2}, title = {Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome}, url = {http://dx.doi.org/10.1126/science.1117389}, volume = {309}, year = {2005} }

TY - JOUR ID - citeulike:320356 L3 - citeulike-article-id:320356 TI - Accurate Multiplex Polony Sequencing of an Evolved Bacterial Genome JF - Science VL - 309 IS - 5741 SP - 1728 EP - 1732 N2 - 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. KW - genomics KW - sequencing AU - Shendure, Jay AU - Porreca, Gregory AU - Reppas, Nikos AU - Lin, Xiaoxia AU - Mccutcheon, John AU - Rosenbaum, Abraham AU - Wang, Michael AU - Zhang, Kun AU - Mitra, Robi AU - Church, George PY - 2005/09/09/ UR - http://dx.doi.org/10.1126/science.1117389 ER -