Ultra-high-speed DNA sequencing using capillary electrophoresis chips

Anal Chem. 1995 Oct 15;67(20):3676-80. doi: 10.1021/ac00116a010.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Sequence
  • DNA / analysis
  • DNA / chemistry*
  • DNA Primers / chemistry
  • Electrophoresis, Capillary / instrumentation*
  • Electrophoresis, Polyacrylamide Gel
  • Glass
  • Molecular Sequence Data
  • Sequence Analysis, DNA / methods*

Substances

  • DNA Primers
  • DNA