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G Singh, WW Hauswirth, WE Ross and AH Neims
This paper describes a rapid and reliable method for quantification of damage to mitochondrial DNA (mtDNA), especially strand breaks. The degree of damage to mtDNA is assessed by the proportion of physical forms (i.e., supercoiled versus open-circular and linear forms) upon agarose gel electrophoresis, blotting, and visualization by hybridization with [32P]mtDNA probes. The use of a radiolabeled probe is a crucial step in the procedure because it provides both a means to quantify by radioautography and to obtain the mtDNA specificity required to eliminate misinterpretation due to nuclear DNA contamination. To demonstrate the utility of this technique, X- irradiation and epichlorohydrin are shown to damage both isolated mtDNA and mtDNA in whole cells in a dose-dependent fashion.
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J. Chen, F. F. Kadlubar, and J. Z. Chen DNA supercoiling suppresses real-time PCR: a new approach to the quantification of mitochondrial DNA damage and repair Nucleic Acids Res., March 13, 2007; (2007) gkm010v2. [Abstract] [Full Text] [PDF] |
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