O-Hydroxylamine-Coupled Alkaline Gel Electrophoresis Assay for the Detection and Measurement of DNA Single-Strand Breaks
The ability to detect and measure DNA single-strand breaks has been the aim of numerous assays developed to assess genotoxicity. These methods often rely on alkaline conditions to denature DNA. However, alkaline treatment of DNA also introduces artifactual SSBs through the cleavage of alkali-labile sites resulting in confounded data. Here, we describe a modified alkaline gel electrophoresis assay coupled with a neutral O-hydroxylamine to obtain the measurement of true SSB formation.
Key wordsAgarose Alkaline Electrophoresis O-hydroxylamine Single-strand breaks
This work was supported by National Institute of Environmental Health Science grants T32-ES07126, P42-ES05948, and P30-ES10126 and National Cancer Institute grant T32-CA72319.
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