Molecular Biotechnology

, Volume 4, Issue 1, pp 25–43 | Cite as

Preparation, manipulation, and pulse strategy for one-dimensional pulsed-field gel electrophoresis (ODPFGE)

  • Jaan Noolandi
  • Chantal Turmel


The underlying principles for zero-integrated-field electrophoresis (ZIFE) pulses and more general forward-biased pulse schemes are reviewed for one-dimensional pulsed-field gel electrophoresis (ODPFGE) separations of large DNA molecules. Detailed descriptions of materials, preparation protocols, hardware requirements, and procedures are given. A variety of gel pictures for known yeast DNA markers are shown.

Index Entries

One-dimensional pulsed-field gel electrophoresis zero-integrated-field electrophoresis DNA preparation DNA manipulation pulse strategy 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Lalande, M., Kunkel, L. M., Flint, A., and Latt, S. A. (1984) Development and use of metaphase chromosome flow sorting methodology to obtain recombinant phage libraries enriched for parts of the human X chromosome.Cytometry 5, 101–107.PubMedCrossRefGoogle Scholar
  2. 2.
    Feinberg, A. P. and Vogelstein, B. (1983) A technique for radio-labelling DNA restriction endonuclease fragments to high specific activity.Anal. Biochem. 132, 6–13.PubMedCrossRefGoogle Scholar
  3. 3.
    Barlow, D. P. (1992) Preparation, restriction and hybridization analyses of mammalian genomic DNA for pulse-field gel electrophoresis, inMethods in Molecular Biology, vol. 12. Pulse Field Gel Electrophoresis (Burmeister, M. and Ulanovsky, L., eds.), Humana, Totowa, NJ, pp. 107–128.CrossRefGoogle Scholar
  4. 4.
    Noolandi, J. (1992) Theory of DNA gel electrophoresis, inAdvances in Electrophoresis, vol. 5 (Chrambach, A., Dunn, M. J., and Radola, B. J., eds.), VCH, New York, pp. 1–57.Google Scholar
  5. 5.
    Lalande, M., Noolandi, J., Turmel, C., Rousseau, J., and Slater, G. W.,(1987) Pulsed-field electrophoresis: Application of a computer model to the separation of large DNA molecules.Proc. Natl. Acad. Sci. USA 84, 8011–8015.PubMedCrossRefGoogle Scholar
  6. 6.
    Slater, G. W., Rousseau, J., Noolandi, J., Turmel, C., and Lalande, J. (1988) Quantitative analysis of the three regimes of DNA electrophoresis in agarose gels.Biopolymers 27, 509–524.PubMedCrossRefGoogle Scholar
  7. 7.
    Turmel, C., Brassard, E., Slater, G. W., and Noolandi, J. (1989) Molecular detrapping and band narrowing with high frequency modulation of pulsed field electrophoresis.Nucleic Acids Res. 18, 569–575.CrossRefGoogle Scholar
  8. 8.
    Olson, M. V. (1989) Pulsed field gel electrophoresis, inGenetic Engineering (Setlow, J. K., ed.), Plenum, New York, pp. 183–227.Google Scholar
  9. 9.
    Heller, C. and Pohl, M. (1990) Field inversion electrophoresis with different pulse time ramps.Nucleic Acids Res. 18, 6299–6304.PubMedCrossRefGoogle Scholar
  10. 10.
    Birren, B. W., Lai, E., Clark, S. M. Hood, L., and Simon, M. I. (1988) Optimized conditions for pulsed field gel electrophoretic separations of DNA.Nucleic Acids Res. 16, 7563–7582.PubMedCrossRefGoogle Scholar
  11. 11.
    Fan, J. B., Chikashige, Y., Smith, C. L., Niwa, O., Yanagida, M., and Cantor, C. R. (1988) Construction of a Not I restriction map of the fission yeastSchizosaccharomyces pombe genome.Nucleic Acids Res. 17, 2801–2818.CrossRefGoogle Scholar
  12. 12.
    Lasker, B. A., Carle, G. F., Kobayashi, G. S., and Medoff, G. (1989) Comparison of the separation ofCandida albicans chromosome-sized DNA by pulsed field gel electrophoresis techniques.Nucleic Acids Res. 17, 3783–3793.PubMedCrossRefGoogle Scholar
  13. 13.
    Chu, G. (1990) Pulsed field gel electrophoresis: Theory and practice.Methods: A Companion to Methods Enzymol. 1, 129–142.CrossRefGoogle Scholar
  14. 14.
    Noolandi, J. and Turmel, C. (1992) Preparation and separation of intact chromosomes of vertebrates by one-dimensional pulsed-field gel electrophoresis (ODPFGE), inMethods in Molecular Biology, vol. 12. Pulse Field Gel Electrophoresis (Burmeister, M. and Ulanovsky, L., eds.), Humana, Totowa, NJ, pp. 135–143.CrossRefGoogle Scholar
  15. 15.
    Higgins, M. J., Turmel, C., Noolandi, J., Neumann, P., and Lalande, M. (1990) Construction of the physical map for three loci in chromosome band 13q14: Comparison to the genetic map.Proc. Natl. Acad. Sci. USA 87, 3415–3419.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc 1995

Authors and Affiliations

  • Jaan Noolandi
    • 1
  • Chantal Turmel
    • 1
  1. 1.Xerox Research Centre of CanadaMississaugaCanada

Personalised recommendations