Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis

John Clark Sutherland; Kiley J. Reynolds; David J. Fisk

doi:10.1117/12.237621

1 April 1996 Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis

John Clark Sutherland, Kiley J. Reynolds, David J. Fisk

Proceedings Volume 2680, Ultrasensitive Biochemical Diagnostics; (1996) https://doi.org/10.1117/12.237621
Event: Photonics West '96, 1996, San Jose, CA, United States

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.

Citation Download Citation

John Clark Sutherland, Kiley J. Reynolds, and David J. Fisk "Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); https://doi.org/10.1117/12.237621

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