Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing.
An efficient, nanoliter-scale microfabricated bioprocessor integrating all three Sanger sequencing steps, thermal cycling, sample purification, and capillary electrophoresis, has been developed and evaluated. Hybrid 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 complex genomes. The performance of this miniaturized DNA sequencer provides a benchmark for predicting the ultimate cost and efficiency limits of Sanger sequencing.[1]References
- Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing. Blazej, R.G., Kumaresan, P., Mathies, R.A. Proc. Natl. Acad. Sci. U.S.A. (2006) [Pubmed]
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