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The estimation of oxidative DNA damage using the single-cell gel electrophoresis (commet assay).
Škubalová, Věra ; Hochmann, Jiří (advisor) ; Slyšková, Jana (referee)
The oxidative cell damage represents one of the most common type of damage, which concerns all cell components. It produces the oxidative modifications of lipid structure, proteins and DNA. We concentrated on the last one in this work. The oxidative changes result in modification of replication cell cycle, which are represented for example by carcinogenesis, mutagenesis or by cell aging. Cells are provided by repairing mechanism. Owing to this mechanism the cells are able to repair this damage and stay viable. The oxidative damage is induced by various factors, for example free oxygen radicals or UV radiation, which we used for producing oxidative damage of DNA in this work. It is possible to measure the oxidative damage by using different methods, the most common way is using the comet assay. It represents an important method used for detection of single and doublestrand breaks, for detection of alcali-labile sites, searching for DNA- DNA/DNAprotein cross-linking, oxidative and alkylate damage of DNA. Only a little number of cells are needed for the measurement, which is the most important advantage of this method. We also used the modification of comet assay. It means the application of the reparative enzymes, which are able to recognize the specific oxidative damage of DNA. We used three...
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The estimation of oxidative DNA damage using the single-cell gel electrophoresis (commet assay).
Škubalová, Věra ; Hochmann, Jiří (advisor) ; Slyšková, Jana (referee)
The oxidative cell damage represents one of the most common type of damage, which concerns all cell components. It produces the oxidative modifications of lipid structure, proteins and DNA. We concentrated on the last one in this work. The oxidative changes result in modification of replication cell cycle, which are represented for example by carcinogenesis, mutagenesis or by cell aging. Cells are provided by repairing mechanism. Owing to this mechanism the cells are able to repair this damage and stay viable. The oxidative damage is induced by various factors, for example free oxygen radicals or UV radiation, which we used for producing oxidative damage of DNA in this work. It is possible to measure the oxidative damage by using different methods, the most common way is using the comet assay. It represents an important method used for detection of single and doublestrand breaks, for detection of alcali-labile sites, searching for DNA- DNA/DNAprotein cross-linking, oxidative and alkylate damage of DNA. Only a little number of cells are needed for the measurement, which is the most important advantage of this method. We also used the modification of comet assay. It means the application of the reparative enzymes, which are able to recognize the specific oxidative damage of DNA. We used three...
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