Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis John C. Sutherland, Kiley J. Reynolds and David, J. Fisk SPIE 2680, 326-340 (1996). 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 electrophoresis 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.

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