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.
Blazej
Robert G
RG
University of California, San Francisco/University of California, Berkeley, Joint Bioengineering Graduate Group, Berkeley, CA 94720, USA.
Kumaresan
Palani
P
Mathies
Richard A
RA
D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004247:DNA / Q000235:genetics*; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods*
D004247:DNA