Agarose Gel Electrophoresis for DNA Damage Analysis
Physical and chemical mutagens induce frank breaks in DNA which reduce its single-strand molecular weight. Other nonbreak lesions in the DNA can often be converted into strand breaks by chemical and enzymatic means. Using agarose gel electrophoresis along with various cleavage schemes, the average density of breaks and various lesion classes along mammalian DNA can be determined.
KeywordsAcridine Orange Acridine Orange Abasic Site Lesion Class Pyrimidone Photoproduct
Unable to display preview. Download preview PDF.
- Drouin, R., Rodriguez, H., Gao, S., Gebreyes, Z., O’Connor, T. R., Holmquist, G. P., and Akman, S. A. (1996). Cupric ion/ascorbate/hydrogen peroxide-induced DNA damage: DNA-bound copper ion primarily induces base modifications. Free Radical Biol. Med. (in press).Google Scholar
- Holmquist, G. P. (1988). DNA sequences in G-bands and R-bands, in:Chromosomes and Chromatin (K. W. Adolph, ed.), CRC Press, Boca Raton, FL, pp. 75–121.Google Scholar
- Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
- Pfeifer, G. P., Drouin, R., Riggs, A. D., and Holmquist, G. P. (1991). In vivo mapping of a DNA adduct at nucleotide resolution: Detection of pyrimidine (6−4) pyrimidone photoproducts by ligation-mediated polymerase chain reaction. Proc. Natl. Acad. Sci. USA 88:1374–1378.PubMedCrossRefGoogle Scholar
- Rigler, R. (1966). Microfluorometric characterization of intracellular nucleic acids and nucleoproteins by acridine orange. Acta Physiol. Scand. 67(Suppl. 267):1–122.Google Scholar
- Rodriguez, H., Drouin, R., Holmquist, G. P., O’Connor, T. R., Boiteux, S., Laval, J., Doroshow, J. H., and Akman, S. A. (1995). Mapping of copper/hydrogen peroxide-induced DNA damage at nucleotide resolution in human genomic DNA by ligation-mediated PCR. J. Biol. Chem. 270:17633–17640.PubMedCrossRefGoogle Scholar
- Sambrook, J., Fritsch, E. F, and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar
- Tanford, C. (1961). Physical Chemistry of Macromolecules, Wiley, New York.Google Scholar