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

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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 ... continued below

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15 p.

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Sutherland, J.C.; Reynolds, K.J. & Fisk, D.J. April 1, 1996.

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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.

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15 p.

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OSTI as DE96008872

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  • Photonics West `96: conference on quantum well and superlattice physics VI, San Jose, CA (United States), 27 Jan - 2 Feb 1996

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  • Other: DE96008872
  • Report No.: BNL--62891
  • Report No.: CONF-960163--14
  • Grant Number: AC02-76CH00016
  • Office of Scientific & Technical Information Report Number: 207500
  • Archival Resource Key: ark:/67531/metadc669257

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  • April 1, 1996

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  • June 29, 2015, 9:42 p.m.

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  • Dec. 11, 2015, 3:20 p.m.

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Sutherland, J.C.; Reynolds, K.J. & Fisk, D.J. Dispersion functions and factors that determine resolution for DNA sequencing by gel electrophoresis, article, April 1, 1996; Upton, New York. (https://digital.library.unt.edu/ark:/67531/metadc669257/: accessed April 23, 2019), University of North Texas Libraries, Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.