A direct view by immunofluorescent comet assay (IFCA) of DNA damage induced by nicking and cutting enzymes, ionizing (137)Cs radiation, UV-A laser microbeam irradiation and the radiomimetic drug bleomycin

Mutagenesis
Paulius GrigaraviciusKarl Otto Greulich

Abstract

In DNA repair research, DNA damage is induced by different agents, depending on the technical facilities of the investigating researchers. A quantitative comparison of different investigations is therefore often difficult. By using a modified variant of the neutral comet assay, where the histone H1 is detected by immunofluorescence [immunofluorescent comet assay (IFCA)], we achieve previously unprecedented resolution in the detection of fragmented chromatin and show that trillions of ultraviolet A photons (of a few eV), billions of bleomycin (BLM) molecules and thousands of gamma quanta (of 662 keV) generate, in first order, similar damage in the chromatin of HeLa cells. A somewhat more detailed inspection shows that the damage caused by 20 Gy ionizing radiation and by a single laser pulse of 10 microJ are comparable, while the damage caused by 12 microg/ml BLM depends highly on the individual cell. Taken together, this work provides a detailed view of DNA fragmentation induced by different treatments and allows comparing them to some extent, especially with respect to the neutral comet assay.

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Feb 20, 2008·Mutagenesis·Andrew R CollinsRudolf Stetina

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Citations

Nov 23, 2017·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Rodrigo C BohrerVilceu Bordignon
Jul 20, 2010·Analytical and Bioanalytical Chemistry·Hanna L Karlsson
Nov 9, 2018·Radiation Research·J SchuemannS J McMahon
Dec 23, 2016·Reports on Progress in Physics·Karl Otto Greulich

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