PL EN


Preferences help
enabled [disable] Abstract
Number of results
2000 | 47 | 3 | 835-839
Article title

Densitometric quantification of DNA topoisomers in ethidium bromide-stained agarose gels and chemiluminescence-detected X ray films.

Authors
Content
Title variants
Languages of publication
EN
Abstracts
EN
This study investigates the contribution of deformational strain imposed by topological interconversions of DNA in ethidium bromide-binding on agarose gels. Closed-circular plasmid DNAs were nicked using UV exposure and the DNA bands were quantified by densitometry. The results show that the closed circular DNA binds the same amount of the dye as its nicked counterpart. The relationship between the band intensity on X-ray films of chemiluminescence-detected Southern blots and DNA concentration was shown to be linear.
Publisher

Year
Volume
47
Issue
3
Pages
835-839
Physical description
Dates
published
2000
received
2000-05-29
accepted
2000-06-12
Contributors
author
  • Department of Pharmaceutical Biotechnology, School of Pharmacy, Ege University, Izmir, Turkey
References
  • 1. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.M., Seidman, J.G. & Struhl, K. (1987) Current Protocols in Molecular Biology; pp. 2.5.1-2.5.9, John Wiley and Sons Inc., New York.
  • 2. Heath, P.J., Clendenning, J.B., Fujimoto, B.S. & Schurr, J.M. (1996) Effect of bending strain on the torsion elastic constant of DNA. J. Mol. Biol. 260, 718-730.
  • 3. Holtke, H.J., Ankenbauer, W., Muehleg, K., Rein, R., Sagner, G., Seibl, R. & Walter, T. (1995) The digoxigenin (DIG) system for non-radioactive labelling and detection of nucleic acids An overview. Cell. Mol. Biol. 41, 883-905.
  • 4. Lazarus, G.M., Henrich, J.P., Kelly, W.G., Schmitz, S.A. & Castora, F.J. (1987) Purification and characteristics of a type I topoisomerase from calf thymus mitochondria. Arch. Biochem. Biophys. 293, 201-207.
  • 5. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248-254.
  • 6. Topcu, Z. & Castora, F.J. (1995) Mammalian mitochondrial DNA topoisomerase I preferentially relaxes supercoils in plasmids containing specific mitochondrial DNA sequences. Biochim. Biophys. Acta 1264, 377-387.
  • 7. Bauer, W. & Vinograd, J. (1970) Interaction of closed circular DNA with intercalative dyes; the free energy of superhelix formation. J. Mol. Biol. 47, 419-435.
  • 8. Oppenheim, A. (1981) Separation of closed circular DNA from linear DNA by electrophoresis in two dimensions in agarose gels. Nucleic Acids Res. 9, 6805-6812.
  • 9. Bates, A.D. & Maxwell, A. (1993) DNA Supercoiling; in DNA Topology (Rickwood, D., ed) pp. 17-45, IRL-Press Inc, New York.
Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv47i3p835kz
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.