학술논문
Genome sequencing in microfabricated high-density picolitre reactors.
Document Type
Article
Author
Margulies, Marcel; Egholm, Michael; Altman, William E.; Attiya, Said; Bader, Joel S.; Bemben, Lisa A.; Berka, Jan; Braverman, Michael S.; Yi-Ju Chen; Zhoutao Chen; Dewell, Scott B.; Lei Du; Fierro, Joseph M.; Gomes, Xavier V.; Godwin, Brian C.; Wen He; Helgesen, Scott; Chun He Ho; Irzyk, Gerard P.; Jando, Szilveszter C.
Source
Subject
*GENOMES
*GENETICS
*GENOMICS
*DNA
*NUCLEIC acids
*
*
*
*
Language
ISSN
0028-0836
Abstract
The proliferation of large-scale DNA-sequencing projects in recent years has driven a search for alternative methods to reduce time and cost. Here we describe a scalable, highly parallel sequencing system with raw throughput significantly greater than that of state-of-the-art capillary electrophoresis instruments. The apparatus uses a novel fibre-optic slide of individual wells and is able to sequence 25 million bases, at 99% or better accuracy, in one four-hour run. To achieve an approximately 100-fold increase in throughput over current Sanger sequencing technology, we have developed an emulsion method for DNA amplification and an instrument for sequencing by synthesis using a pyrosequencing protocol optimized for solid support and picolitre-scale volumes. Here we show the utility, throughput, accuracy and robustness of this system by shotgun sequencing and de novo assembly of the Mycoplasma genitalium genome with 96% coverage at 99.96% accuracy in one run of the machine. [ABSTRACT FROM AUTHOR]