Genome sequencing in microfabricated high-density picolitre reactors
Abstrak
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.
Penulis (56)
M. Margulies
M. Egholm
William E. Altman
S. Attiya
J. Bader
Lisa A. Bemben
J. Berka
Michael S. Braverman
Yi-Ju Chen
Zhoutao Chen
Scott B. Dewell
Lei Du
J. M. Fierro
Xavier V. Gomes
B. Godwin
Wenshe He
S. Helgesen
Chun He Ho
G. Irzyk
Szilveszter C. Jando
Maria L. I. Alenquer
T. Jarvie
K. Jirage
Jong-Bum Kim
James R. Knight
Janna R. Lanza
J. Leamon
S. Lefkowitz
M. Lei
Jing Li
K. Lohman
Hong Lu
V. Makhijani
K. McDade
M. McKenna
E. Myers
E. Nickerson
J. Nobile
Ramona Plant
Bernard P. Puc
Michael T. Ronan
G. T. Roth
G. Sarkis
J. Simons
J. W. Simpson
Maithreyan Srinivasan
K. Tartaro
A. Tomasz
K. Vogt
G. Volkmer
Shally H. Wang
Yong Wang
M. Weiner
Pengguang Yu
R. Begley
J. Rothberg
Akses Cepat
- Tahun Terbit
- 2005
- Bahasa
- en
- Total Sitasi
- 7987×
- Sumber Database
- Semantic Scholar
- DOI
- 10.1038/nature03959
- Akses
- Open Access ✓