Ultra-high-speed DNA sequencing using capillary electrophoresis chips.
DNA sequencing has been performed on microfabricated capillary electrophoresis chips. DNA separations were achieved in 50 x 8 microns cross-section channels microfabricated in a 2 in. x 3 in. glass sandwich structure using a denaturing 9% T, 0% C polyacrylamide sieving medium. DNA sequencing fragment ladders were produced and fluorescently labeled using the recently developed energy transfer dye-labeled primers. Sequencing extension fragments were separated to approximately 433 bases in only 10 min using a one-color detection system and an effective separation distance of only 3.5 cm. Using a four-color labeling and detection format, DNA sequencing with 97% accuracy and single-base resolution to approximately 150 bases was achieved in only 540 s. A resolution of greater than 0.5 was obtained out to 200 bases for both the one- and four-color separations. The prospects for enhancing the resolution and sensitivity of these chip separations are discussed. This work establishes the feasibility of high-speed, high-throughput DNA sequencing using capillary array electrophoresis chips.
Woolley
A T
AT
Department of Chemistry, University of California, Berkeley 94720, USA.
Mathies
R A
RA
D001483:Base Sequence; D004247:DNA / Q000032:analysis / Q000737:chemistry*; D017931:DNA Primers / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004591:Electrophoresis, Polyacrylamide Gel; D005898:Glass; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*
D017931:DNA Primers; D004247:DNA