Microfabricated bioprocessor for integrated nanoliter-scale Sanger DNA sequencing

摘要

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.