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Nucleic Acids Research, Vol 27, Issue 24 e36-e36, Copyright © 1999 by Oxford University Press

ARTICLES

A multiple-capillary electrophoresis system for small-scale DNA sequencing and analysis

J Zhang, KO Voss, DF Shaw, KP Roos, DF Lewis, J Yan, R Jiang, H Ren, JY Hou, Y Fang, X Puyang, H Ahmadzadeh and NJ Dovichi
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada

A five-capillary system has been developed for DNA sequencing and analysis. The post-column fluorescence detector is based on a sheath-flow cuvette. The instrument provides uniform and continuous illumination of the samples. The cuvette virtually eliminates cross-talk in the fluorescence signal between capillaries. Discrete single-photon counting avalanche photodiodes provide high efficiency light detection. The instrument has detection limits (3[sigma]) of 130 +- 30 fluorescein molecules injectedonto each capillary. Over 650 bases of sequence at 98.8% accuracy were generated in 100 min at 50[deg]C from M13mp18. Separation and detection of short tandem repeats proved efficient and accurate with theuse of internal standards for direct comparison of migration times between capillaries.
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