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Title: Ultra-high-speed DNA sequencing using capillary electrophoresis chips

Abstract

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.

Authors:
;  [1]
  1. Univ. of California, Berkeley, CA (United States)
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
137384
Resource Type:
Journal Article
Journal Name:
Analytical Chemistry (Washington)
Additional Journal Information:
Journal Volume: 67; Journal Issue: 20; Other Information: PBD: 15 Oct 1995
Country of Publication:
United States
Language:
English
Subject:
40 CHEMISTRY; 55 BIOLOGY AND MEDICINE, BASIC STUDIES; 42 ENGINEERING NOT INCLUDED IN OTHER CATEGORIES; 66 PHYSICS; DNA; QUANTITATIVE CHEMICAL ANALYSIS; DNA SEQUENCING; ELECTROPHORESIS; MICROPROCESSORS

Citation Formats

Woolley, A T, and Mathies, R A. Ultra-high-speed DNA sequencing using capillary electrophoresis chips. United States: N. p., 1995. Web. doi:10.1021/ac00116a010.
Woolley, A T, & Mathies, R A. Ultra-high-speed DNA sequencing using capillary electrophoresis chips. United States. https://doi.org/10.1021/ac00116a010
Woolley, A T, and Mathies, R A. 1995. "Ultra-high-speed DNA sequencing using capillary electrophoresis chips". United States. https://doi.org/10.1021/ac00116a010.
@article{osti_137384,
title = {Ultra-high-speed DNA sequencing using capillary electrophoresis chips},
author = {Woolley, A T and Mathies, R A},
abstractNote = {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.},
doi = {10.1021/ac00116a010},
url = {https://www.osti.gov/biblio/137384}, journal = {Analytical Chemistry (Washington)},
number = 20,
volume = 67,
place = {United States},
year = {1995},
month = {10}
}