Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 103, Issue 19, Pages 7240-7245Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0602476103
Keywords
capillary electrophoresis; genetic analysis; microfluidic
Categories
Funding
- NHGRI NIH HHS [R01 HG001399, HG00047, R01 HG003583, HG01399, T32 HG000047, K22 HG000047, HG003583] Funding Source: Medline
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An efficient, nanc liter-scale microfabricated bioprocessor integrating all three Sar ger sequencing steps, thermal cycling, sample purification, and capillary electrophoresis, has been developed and evaluated. Hybric glass-polydimethylsiloxane (PDMS) wafer-scale construction is used to combine 250-nl reactors, affinity-capture purification chambers, high-performance capillary electrophoresis channels, and pneumatic valves and pumps onto a single microfabricated device. Lab-on-a-chip-level integration enables complete Sanger sequencing from only 1 fmol of DNA template. Up to 556 continuous bases were sequenced with 99% accuracy, demonstrating read lengths required for de novo sequencing of human and other complete (K) genomes. The performance of this miniaturized DNA sequencer provides a benchmark for predicting the ultimate cost and efficiency limits of Sanger sequencing.
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