| Автор | Aborn, James H. |
| Автор | El-Difrawy, Sameh A. |
| Автор | Novotny, Mark |
| Автор | Gismondi, Elizabeth A. |
| Автор | Lam, Roger |
| Автор | Matsudaira, Paul |
| Автор | Mckenna, Brian K. |
| Автор | O'Neil, Thomas |
| Автор | Streechon, Philip |
| Автор | Ehrlich, Daniel J. |
| Дата выпуска | 2005 |
| dc.description | A 768-lane DNA sequencing system based on microfluidic plates has been designed as a near-term successor to 96-lane capillary arrays. Electrophoretic separations are implemented for the first time in large-format (25 cm à 50 cm) microdevices, with the objective of proving realistic read length, parallelism, and the scaled sample requirements for long-read de novo sequencing. Two 384-lane plates are alternatively cycled between electrophoresis and regeneration via a robotic pipettor. A total of greater than 172â 000 bases, 99% accuracy (corresponding to quality score 20) is achieved for each iteration of a 384 lane plate. At current operating conditions, this implies a system throughput exceeding 4 megabases of raw sequence (Phred 20) per day on the new platform. Standard operation is at â 1/32à â Sanger chemistry, equal to typical genome center operation on mature capillary array machines, and a 16-fold improvement in scaling relative to previous microfabricated devices. Experiments provide evidence that sample concentration can be further reduced to 1/256à Sanger chemistry in the microdevice. Life-testing indicates a usable life of >150 hours (more than 50 runs) for the 384 lane plates. The combined advances, particularly those in read length and sample requirement, directly address the cost model requirements for adaptation of the new technology as the next step beyond capillary array instruments. |
| Формат | application.pdf |
| Издатель | Royal Society of Chemistry |
| Название | A 768-lane microfabricated system for high-throughput DNA sequencing |
| Тип | research-article |
| DOI | 10.1039/b501104c |
| Electronic ISSN | 1473-0189 |
| Print ISSN | 1473-0197 |
| Журнал | Lab on a Chip |
| Том | 5 |
| Первая страница | 669 |
| Последняя страница | 674 |
| Аффилиация | Aborn James H.; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | El-Difrawy Sameh A.; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Novotny Mark; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Gismondi Elizabeth A.; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Lam Roger; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Matsudaira Paul; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Mckenna Brian K.; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | O'Neil Thomas; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Streechon Philip; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Аффилиация | Ehrlich Daniel J.; Whitehead Institute for Biomedical Research, Nine Cambridge Center |
| Выпуск | 6 |
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