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

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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsGerald E. Cohn, Steven A. Soper, C.H.W. Chen
Pages326-340
Number of pages15
StatePublished - 1996
Externally publishedYes
EventUltrasensitive Biochemical Diagnostics - San Jose, CA, USA
Duration: Jan 31 1996Feb 2 1996

Publication series

NameProceedings of SPIE - The International Society for Optical Engineering
Volume2680
ISSN (Print)0277-786X

Other

OtherUltrasensitive Biochemical Diagnostics
CitySan Jose, CA, USA
Period1/31/962/2/96

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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