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Proceedings Paper

Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis
Author(s): John Clark Sutherland; Kiley J. Reynolds; David J. Fisk
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Paper Abstract

The number of bases that can be read in a single run by a DNA sequencing instrument that detects fluorophore labeled DNA arriving at a 'finish-line' located a fixed distance from the starting wells is influenced by numerous parameters. Strategies for improving the length-of- read of a DNA sequencer can be based on quantitative models of the separation of DNA by gel electrophoresis. The dispersion function of the electrophoretic system -- the relationship between molecular contour length and time of arrival at the detector -- is useful in characterizing the performance of a DNA sequencer. We adapted analytical representations of dispersion functions, originally developed for snapshot imaging of DNA gels, (samples electrophoresed for constant time), to finish-line imaging, and demonstrated that a logistic- type function with non-integral exponent is required to describe the experimental data. We use this dispersion function to determine the resolution length and resolving power of a LI-COR DNA sequencing system and a custom built capillary gel electrophoresis system, and discuss the factors that presently limit the number of bases that can be determined reliably in a single sequencing run.

Paper Details

Date Published: 1 April 1996
PDF: 15 pages
Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); doi: 10.1117/12.237621
Show Author Affiliations
John Clark Sutherland, Brookhaven National Lab. (United States)
Kiley J. Reynolds, Brookhaven National Lab. (United States)
David J. Fisk, Brookhaven National Lab. (United States)

Published in SPIE Proceedings Vol. 2680:
Ultrasensitive Biochemical Diagnostics
Gerald E. Cohn; Steven A. Soper; C. H. Winston Chen, Editor(s)

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