2005/08/04 by Jay Shendure, Gregory J. Porreca, Nikos B. Reppas +7 · 3 citations
Biochemistry, Genetics and Molecular Biology · Environmental Science · #Genomics and Phylogenetic Studies #Bacteriophages and microbial interactions #RNA and protein synthesis mechanisms #Multiplex #DNA sequencing #Biology #Computational biology #DNA #Polymerase chain reaction #Multiplex polymerase chain reaction #Genetics #Gene
paper · doi:10.1126/science.1117389
openalex publication_date 2005/08/04 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/23
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.