Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis
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.
- Research Organization:
- Brookhaven National Lab. (BNL), Upton, NY (United States)
- Sponsoring Organization:
- USDOE, Washington, DC (United States)
- DOE Contract Number:
- AC02-76CH00016
- OSTI ID:
- 207500
- Report Number(s):
- BNL-62891; CONF-960163-14; ON: DE96008872
- Resource Relation:
- Conference: Photonics West `96: conference on quantum well and superlattice physics VI, San Jose, CA (United States), 27 Jan - 2 Feb 1996; Other Information: PBD: [1996]
- Country of Publication:
- United States
- Language:
- English
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