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A Modified AlkalineComet Assay forMeasuring DNA RepairCapacity in HumanPopulations Andrzej TrzeciakJanice BarnesMichele K. Evans2007 год

A Modified Alkaline Comet Assay for Measuring DNA Repair Capacity in Human Populations
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Аннотация: Trzeciak, A. R., Barnes, J. and Evans, M. K. A Modified Alkaline Comet Assay for Measuring DNA Repair Capacity in Human Populations. Radiat. Res. 169, 110-121 (2008). Use of the alkaline comet assay to assess DNA repair capacity in human populations has been limited by several factors, including lack of methodology for use of unstimulated cryopreserved peripheral blood mononuclear cells (PBMCs), insufficient control of interexperimental variability, and limited analysis of DNA repair kinetics. We show that unstimulated cryopreserved PBMCs can be used in DNA repair studies performed using the comet assay. We have applied data standardization for the analysis of DNA repair capacity using negative and positive internal standards as controls for interexperimental variability. Our standardization procedure also uses negative controls, which provides a way to minimize the interference of interindividual variation in baseline DNA damage levels on DNA repair capacity measurements in populations. DNA repair capacity was assessed in a small human cohort using the parameters described in the literature including initial DNA damage, half-time of DNA repair, and residual DNA damage after 30 and 60 min. We have also introduced new DNA repair capacity parameter, initial rate of DNA repair. There was no difference in DNA repair capacity between fresh and cryopreserved PBMCs when measured by the Olive tail moment and tail DNA. The use of DNA repair capacity parameters in assessment of fast and slow single-strand break repair components is discussed.
Год издания: 2007
Авторы: Andrzej Trzeciak, Janice Barnes, Michele K. Evans
Издательство: Radiation Research Society
Источник: Radiation Research
Ключевые слова: Carcinogens and Genotoxicity Assessment, DNA Repair Mechanisms, Effects and risks of endocrine disrupting chemicals
Другие ссылки: Radiation Research (HTML)
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PubMed Central (HTML)
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