Genome sequencing in microfabricated high-density picolitre reactors.
Nature
; 437(7057): 376-80, 2005 Sep 15.
Article
em En
| MEDLINE
| ID: mdl-16056220
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Genoma Bacteriano
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Análise de Sequência de DNA
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Genômica
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Mycoplasma genitalium
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Microquímica
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article