Twelve-gel slide format optimised for comet assay and fluorescent in situ hybridisation

• DOI: 10.1016/j.toxlet.2010.02.017
• Publication Date: 2010-02-26T05:37:38Z
• Authors: Sergey Shaposhnikov; Amaya Azqueta; Sara Henriksson; Silja Meier; Isabel Gaivão; Neville H. Huskisson; Andrew Smart; Gunnar Brunborg; Mats Nilsson; Andrew R. Collins
• Keywords: 0301 basic medicine; 03 medical and health sciences; Photosensitizing Agents; Pyrrolidines; Humans; Comet Assay; In Situ Hybridization, Fluorescence; Quinolizines; DNA Damage; HeLa Cells

The comet assay is widely used to measure DNA damage and repair in basic research, genotoxicity testing and human biomonitoring. The conventional format has 1 or 2 gels on a microscope slide, 1 sample per slide. To increase throughput, we have designed and tested a system with 12 smaller gels on one slide, allowing incubation of individual gels with different reagents or enzymes. Thus several times more samples can be analysed with one electrophoresis run, and fewer cells and smaller volumes of test solutions are required. Applications of the modified method include treatment with genotoxic agents at different concentrations; simultaneous analysis of different lesions using a range of enzymes; analysis of cell extracts for DNA repair activity; and fluorescent in situ hybridisation (FISH) to comet DNA with specific labelled probes.