Статья

Genome sequencing in microfabricated high-density picolitre reactors

Marcel Margulies454 Life Sciences Corp., 20 Commercial Street, Branford, Connecticut 06405, USAMichael D. Miller454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAWilliam E. Altman454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USASaid Attiya454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJoel S. Bader454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USALisa A. Bemben454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJan Berka454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAMichael Braverman454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAYi‐Ju Chen454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAZhoutao Chen454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAScott Dewell454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USALei Du454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJoseph M. Fierro454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAXavier V. Gomes454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USABailey Godwin454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAWen He454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAScott Helgesen454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAChun He Ho454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAGerard P. Irzyk454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USASzilveszter C. Jando454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAMaria L. I. Alenquer454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAThomas Jarvie454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAKshama B. Jirage454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJong‐Bum Kim454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJames R. Knight454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJanna R. Lanza454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJohn H. Leamon454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USASteven M. Lefkowitz454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAMing Lei454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJing Li454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAKenton L. Lohman454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAHong Lü454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAVinod B. Makhijani454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAKeith E. McDade454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAMichael P. McKenna454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAEugene W. MyersUniversity of California, California, 94720, Berkeley, USAElizabeth Nickerson454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJohn R. Nobile454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USARamona Plant454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USABernard P. Puc454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAM. T. Ronan454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAGeorge T. Roth454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAGary J. Sarkis454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJan Fredrik Simons454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJohn Simpson454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAMaithreyan Srinivasan454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAKarrie Tartaro454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAAlexander TomaszLaboratory of Microbiology, The Rockefeller University, New York, 10021, New York, USAKari A. Vogt454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAGreg A. Volkmer454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAShally H. Wang454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAYong Wang454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAMichael P. WeinerThe Rothberg Institute for Childhood Diseases, 530 Whitfield Street, Connecticut, 06437, Guilford, USAPengguang Yu454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAR. F. Begley454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USAJonathan M. Rothberg454 Life Sciences Corp., 20 Commercial Street, Connecticut, 06405, Branford, USA

2005en

ABI

Аннотация

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. The race is on for a big prize: the job of providing the world's DNA sequencing laboratories with the successor to the ‘Sanger-based’ technology that gave us the first wave of genome sequences. One technology in the frame is that produced by 454 Life Sciences Corporation of Branford, Connecticut. Today's technology reads 67,000 base pairs per hour; this new approach is 100 times faster, reading 6 million base pairs per hour. The improved performance results from using picolitre-sized chemical reactors, enhanced light-emitting sequencing chemistries and complex informatics. Further miniaturization of the system is planned. Such leaps in technology may one day make it possible to analyse an individual's genome before designing therapy: the ultimate in personalized medicine.

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Идентификаторы

DOI: 10.1038/nature03959

Цитирования и источники

Цитирований: 2Использованных источников: 0

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