Ultra-high-speed DNA fragment separations using microfabricated capillary array electrophoresis chips
- Univ. of California, Berkeley, CA (United States)
Capillary electrophoresis arrays have been fabricated on planar glass substrates by photolithographic masking and chemical etching techniques. The photolithographically defined channel patterns were etched in a glass substrate, and then capillaries were formed by thermally bonding the etched substrate to a second glass slide. High-resolution electrophoretic separations of {phi}X174 Hae III DNA restriction fragments have been performed with these chips using a hydroxyethyl cellulose sieving matrix in the channels. DNA fragments were fluorescently labeled with dye in the running buffer and detected with a laser-excited, confocal fluorescence system. The effects of variations in the electric field, procedures for injection, and sizes of separation and injection channels (ranging from 30 to 120 {mu}m) have been explored. By use of channels with an effective length of only 3.5 cm, separations of {phi}X174 Hae III DNA fragments from {approx}70 to 1000 bp are complete in only 120 sec. We have also demonstrated high-speed sizing of PCR-amplified HLA-DQ{alpha} alleles. This work establishes methods for high-speed, high-throughput DNA separations on capillary array electrophoresis chips. 29 refs., 6 figs.
- Sponsoring Organization:
- USDOE
- OSTI ID:
- 28954
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 24 Vol. 91; ISSN PNASA6; ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
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