Lesion Measurement in Non-Radioactive DNA by Quantitative Gel Electrophoresis
The important role of DNA damage and repair in homeostasis has been clearly demonstrated by studies involving isolated DNA, procaryotic cells and eukaryotic cells growing in culture. The information obtainable from these model systems, while extremely valuable, can never replace the need for data from intact higher organisms. The gel electrophoresis method developed during the past ten years in our laboratories makes possible the quantitation of UV induced pyrimidine dimers, gamma ray induced single- and double-strand breaks and many other types of lesions in nanogram quantities of DNA. The DNA does not have to be labeled with radionuclides or of a particular conformation, thus facilitating the use of the method in measuring damage levels and repair rates in the DNA of intact organisms -- including man.
KeywordsLength Standard Dispersion Function Micrococcus Luteus Ethidium Bromide Fluorescence Ethidium Fluorescence
Unable to display preview. Download preview PDF.
- Ditchburn, R. W., 1965, Light, vols 1, John Wiley, New York.Google Scholar
- Freeman, S. E., Blackett, A. D., Monteleone, D. C., Setlow, R. B., Sutherland, B. M., and Sutherland, J. C., 1986, Quantitation of radiation-, chemical- or enzyme-induced single strand breaks in nonradioactive DNA by alkaline gel electrophoresis: application to pyrimidine dimers, Analvt. Biochem., 158: 119–129.CrossRefGoogle Scholar
- Lett, J. T., 1981, Measurement of single-strand breaks by sedimentation in alkaline sucrose gradients, In, “DNA Repair, A Laboratory Manual of Research Procedures 1, part B”, E. C. Friedberg, and P. C. Hanawalt, eds., Marcel Dekker, Inc. New York.Google Scholar
- Seawell, P. C., Friedberg, E. C., Ganesan, A. K., and Hanawalt, P. C., 1981, Purification of endonuclease V of bacteriophage T4. in, “DNA Repair, A Laboratory Manual of Research Procedures 1, part B”. E. C. Friedberg and P. C. Hanawalt, eds, Marcel Dekker, Inc. New York.Google Scholar
- Sutherland, J. C., Lin, B., Monteleone, D. C., Mugavero, J., Sutherland, B. M., and Trunk, J. 1987a, Electronic imaging system for direct and rapid quantitation of fluorescence from electrophoretic gels: application to ethidium bromide-Stained DNA, Analyt. Biochem., 163: 446–457.PubMedCrossRefGoogle Scholar
- Sutherland, J. C., Monteleone, D. C., Mugavero, J. H., and Trunk, J., 1987b, Undirectional pulsed-field electrophoresis of single- and double-stranded DNA in agarose gels: analytical expressions relating mobility and molecular length and their application in the measurement of strand breaks, Analyt. Biochem., 162: 511–520.PubMedCrossRefGoogle Scholar
- Sutherland, J. C., Bergman, A. M., Chen, C-Z., Monteleone, D. C., Trunk, J., and Sutherland, B. M., 1988, Measurement of DNA damage using gel electrophoresis and electronic imaging, in: “Electrophoresis ’88”, C. Schafer-Nielsen, ed., VCH Verlagsgesellschaft, Weinheim.Google Scholar
- Sutherland, J. C., 1990, Electronic imaging systems for quantitative electrophoresis of DNA. in: “Non-invasive Techniques in Biology and Medicine”, San Francisco Press, (in the press).Google Scholar
- Woodhead, A. D. and Achey, P., 1979, Photoreactivating enzyme in the blind cave fish, Anoptichthys jordani, Comp. Biochem. Physiol., 63B: 73–76.Google Scholar