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Proc Natl Acad Sci U S A. 2006 May 9;103(19):7240-5. Epub 2006 Apr 28.

Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing.

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  • 1University of California, San Francisco/University of California, Berkeley, Joint Bioengineering Graduate Group, Berkeley, CA 94720, USA.

Abstract

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.

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