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Optimized conditions for pulsed field gel electrophoretic separations of DNA

Birren, Bruce W. and Lai, Eric and Clark, Steven M. and Hood, Leroy and Simon, Melvin I. (1988) Optimized conditions for pulsed field gel electrophoretic separations of DNA. Nucleic Acids Research, 16 (15). pp. 7563-7582. ISSN 0305-1048. http://resolver.caltech.edu/CaltechAUTHORS:20120531-143038049

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Abstract

Quantitative measurement of DNA migration in gel electrophoresis requires precisely controlled homogeneous electric fields. A new electrophoresis system has allowed us to explore several parameters governing DNA migration during homogeneous field pulsed field gel (PFG) electrophoresis. Migration was measured at different switch times, temperatures, agarose concentrations, and voltage gradients. Conditions which increase DNA velocities permit separation over a wider size range, but reduce resolution. We have also varied the angle between the alternating electric fields. Reorientation angles between 105° and 165° give equivalent resolution, despite significant differences in DNA velocity. Separation of DNA fragments from 50 to greater than 7000 kilobases (Kb) can easily be optimized for speed and resolution based on conditions we describe.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1093/nar/16.15.7563DOIUNSPECIFIED
http://nar.oxfordjournals.org/content/16/15/7563.abstractPublisherUNSPECIFIED
Additional Information:© 1988 IRL Press, Limited. Received February 29, 1988; Revised and Accepted June 28, 1988. We would like to thank Doug Vollrath for providing a copy of his manuscript prior to publication, John Carbon for providing s. pombe probes, Charles Spence for software development, Petros Arakelian for technical assistance, and Hiroaki Shizuya for his critical reading of the manuscript. This work was supported by grants from the NSF to L.H. (DMB 85-00298) and from NIH to M.I.S. (GM 34236). B.W.B. is a NIH postdoctoral fellow (GM 10974).
Funders:
Funding AgencyGrant Number
NSFDMB 85-00298
NIHGM 34246
NIH Postdoctoral FellowGM 10974
Record Number:CaltechAUTHORS:20120531-143038049
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20120531-143038049
Official Citation: Bnice W. Birren, Eric Lai, Steven M. Clark, Leroy Hood, and Melvin I. Simon Optimized conditions for pulsed field gel electrophoretic separations of DNA Nucl. Acids Res. (1988) 16(15): 7563-7582 doi:10.1093/nar/16.15.7563
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:31755
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:31 May 2012 22:23
Last Modified:26 Dec 2012 15:17

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