Ultra-high-speed DNA sequencing using capillary electrophoresis chips
- Univ. of California, Berkeley, CA (United States)
DNA sequencing has been performed on microfabricated capillary electrophoresis chips. DNA separations were achieved in 50 x 8 {mu}m 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 nearly 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 nearly 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. 22 refs., 4 figs.
- Sponsoring Organization:
- USDOE
- OSTI ID:
- 137384
- Journal Information:
- Analytical Chemistry (Washington), Journal Name: Analytical Chemistry (Washington) Journal Issue: 20 Vol. 67; ISSN ANCHAM; ISSN 0003-2700
- Country of Publication:
- United States
- Language:
- English
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