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1.
Nature ; 475(7356): 348-52, 2011 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-21776081

RESUMO

The seminal importance of DNA sequencing to the life sciences, biotechnology and medicine has driven the search for more scalable and lower-cost solutions. Here we describe a DNA sequencing technology in which scalable, low-cost semiconductor manufacturing techniques are used to make an integrated circuit able to directly perform non-optical DNA sequencing of genomes. Sequence data are obtained by directly sensing the ions produced by template-directed DNA polymerase synthesis using all-natural nucleotides on this massively parallel semiconductor-sensing device or ion chip. The ion chip contains ion-sensitive, field-effect transistor-based sensors in perfect register with 1.2 million wells, which provide confinement and allow parallel, simultaneous detection of independent sequencing reactions. Use of the most widely used technology for constructing integrated circuits, the complementary metal-oxide semiconductor (CMOS) process, allows for low-cost, large-scale production and scaling of the device to higher densities and larger array sizes. We show the performance of the system by sequencing three bacterial genomes, its robustness and scalability by producing ion chips with up to 10 times as many sensors and sequencing a human genome.


Assuntos
Genoma Bacteriano/genética , Genoma Humano/genética , Genômica/instrumentação , Genômica/métodos , Semicondutores , Análise de Sequência de DNA/instrumentação , Análise de Sequência de DNA/métodos , Escherichia coli/genética , Humanos , Luz , Masculino , Rodopseudomonas/genética , Vibrio/genética
2.
Nature ; 437(7057): 376-80, 2005 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-16056220

RESUMO

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.


Assuntos
Genoma Bacteriano , Genômica/instrumentação , Microquímica/instrumentação , Mycoplasma genitalium/genética , Análise de Sequência de DNA/instrumentação , Eletroforese Capilar , Emulsões , Tecnologia de Fibra Óptica , Genômica/economia , Microquímica/economia , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA/economia , Fatores de Tempo
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