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Accurate multiplex polony sequencing of an evolved bacterial genome.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Shendure, Jay Porreca, Gregory Reppas, Nikos B. Lin, Xiaoxia McCutcheon, John P. Rosenbaum, Abraham M. Wang, Michael D. Zhang, Kun Qiu Mitra, Robi David Church, George M. |
| Copyright Year | 2005 |
| 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. |
| Starting Page | 8701 |
| Ending Page | 8706 |
| Page Count | 6 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://arep.med.harvard.edu/pdf/Shendure05b.pdf |
| Alternate Webpage(s) | http://selab.janelia.org/publications/Shendure05/Shendure05-reprint.pdf |
| PubMed reference number | 16081699v1 |
| Volume Number | 309 |
| Issue Number | 5741 |
| Journal | Science |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Base Bead Dosage Form Biopolymer Sequencing Emulsions Genome, Bacterial Instrumentation (attribute) Ligation Microscope Device Component Polymerase Chain Reaction Protocols documentation Reagents part per million (ppm) polyacrylamide gels |
| Content Type | Text |
| Resource Type | Article |
