Pulsed homogeneous orthogonal field gel electrophoresis (PHOGE)
- 1 January 1988
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 16 (15), 7405-7418
- https://doi.org/10.1093/nar/16.15.7405
Abstract
A versatile system (PHOGE) has been developed that allows resolution of molecules of DNA of megabase pair size by the use of homogeneous, orthogonal, pulsed fields. The resulting electrophoretograms have characteristics that differ from those produced by other systems for pulsed field electrophoresis. Molecules in a two-fold range of sizes can be separated with maximum resolution, or a much larger range of sizes may be separated with lower resolution but with a linear relationship of mobility to size from 50 kb, or below, to at least 1 Mb. Straight lanes and large useable gel areas, characteristic of PHOGE, are also valuable for mapping procedures or for any other circumstance in which large numbers of samples of DNA are to be directly compared. Existing models cannot explain the results obtained, because a stage of the molecular reorientation appears to result in a rate of migration greater than that occurring by reptation. We suggest a mechanism that might account for the resolution observed and also suggest that the resolution achieved by existing OFAGE-type systems may be the result of the superimposition of PHOGE and FIGE separatory mechanisms. No maximum size of molecules that may be resolved by the PHOGE system has yet been determined.Keywords
This publication has 11 references indexed in Scilit:
- Theoretical Studies of DNA During Gel ElectrophoresisScience, 1988
- Pulsed-field electrophoresis: application of a computer model to the separation of large DNA molecules.Proceedings of the National Academy of Sciences, 1987
- A model for the separation of large DNA molecules by crossed field gel electrophoresisNucleic Acids Research, 1987
- Separation of Large DNA Molecules by Contour-Clamped Homogeneous Electric FieldsScience, 1986
- Electrophoretic Separations of Large DNA Molecules by Periodic Inversion of the Electric FieldScience, 1986
- Theory of gel electrophoresis of DNABiopolymers, 1985
- Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresisNucleic Acids Research, 1984
- New Techniques for Purifying Large DNAs and Studying Their Properties and PackagingCold Spring Harbor Symposia on Quantitative Biology, 1983
- Electrophoresis of duplex deoxyribonucleic acid in multiple-concentration agarose gels: fractionation of molecules with molecular weights between 2 .times. 106 and 110 .times. 106Biochemistry, 1980
- Electron microscopic observations on the meiotic karyotype of diploid and tetraploid Saccharomyces cerevisiae.Proceedings of the National Academy of Sciences, 1975