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 Citations (Scopus)

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
Volume2680
StatePublished - Jan 1 1996
Externally publishedYes
EventUltrasensitive Biochemical Diagnostics - San Jose, CA, USA
Duration: Jan 31 1996Feb 2 1996

Other

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

Fingerprint

sequencing
electrophoresis
Electrophoresis
DNA
Gels
deoxyribonucleic acid
gels
Optical resolving power
Imaging techniques
Fluorophores
logistics
time constant
arrivals
Logistics
exponents
Detectors
detectors

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Sutherland, J. C., Reynolds, K. J., & Fisk, D. J. (1996). Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis. In G. E. Cohn, S. A. Soper, & C. H. W. Chen (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 2680, pp. 326-340)

Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis. / Sutherland, John C.; Reynolds, Kiley J.; Fisk, David J.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / Gerald E. Cohn; Steven A. Soper; C.H.W. Chen. Vol. 2680 1996. p. 326-340.

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

Sutherland, JC, Reynolds, KJ & Fisk, DJ 1996, Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis. in GE Cohn, SA Soper & CHW Chen (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 2680, pp. 326-340, Ultrasensitive Biochemical Diagnostics, San Jose, CA, USA, 1/31/96.
Sutherland JC, Reynolds KJ, Fisk DJ. Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis. In Cohn GE, Soper SA, Chen CHW, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 2680. 1996. p. 326-340
Sutherland, John C. ; Reynolds, Kiley J. ; Fisk, David J. / Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis. Proceedings of SPIE - The International Society for Optical Engineering. editor / Gerald E. Cohn ; Steven A. Soper ; C.H.W. Chen. Vol. 2680 1996. pp. 326-340
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