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An in vitro approach to study chromosomal DNA damage
Authors:Bala  Madhu  Mathew  Lazar
Affiliation:(1) Department of Molecular Radiation Biology, Institute of Nuclear Medicine and Allied Sciences, Brig. S. K. Mazumdar Marg, 110054, India
Abstract:In this study a simple electrophoresis approach has been proposed for assessing DNA damage per chromosome in vitro. Novel procedures of gel casting, sample loading, electrophoresis and quantification of damage have been suggested. Sets of Saccharomyces cerevisiae chromosomes subjected to DNA damage by Bleomycin, Co60-gamma-radiation alone and in combination with Hoechst were studied in detail. Statistical analyses showed that damage induced by Bleomycin bore linear positive correlation with %GA (r=0.97) and %GT (r=0.61) contents of chromosomes. Samples pre-treated with Hoechst showed much less damage by Co60-gamma-irradiation as compared to samples not treated with Hoechst but exposed to Co60-gamma-irradiation. The `protective effect of Hoechst' bore linear positive correlation (r=0.8) with %TAT content of chromosomes.
Keywords:bleomycin  chromosomal DNA damage  gel electrophoresis  Hoechst  radiation  Saccharomyces cerevisiae
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