Abstract
The comet assay (single cell gel electrophoresis) is a sensitive, versatile method for detecting DNA damage in eukaryotic cells. The traditional comet assay format has 1 or 2 gels on a microscope slide, 1 sample per slide, and there is a limit of 40 gels per experiment given the size of a typical electrophoresis tank. To increase throughput, we have designed and tested a system with 12 minigels on one slide, allowing analysis of up to 12 times more samples in one electrophoresis run. The novel comet assay format compares well with the traditional technology. The various steps are suitable for further automation, and the formats can be adapted to fully automated scoring. The new procedures save time at all stages as fewer slides are handled, and the amounts of reagents needed are reduced significantly. This format is particularly useful for testing of numerous genotoxic agents and nanomaterials at different concentrations and on different types of cells; simultaneous analysis of different lesions using a range of enzymes; and analysis of cell extracts for DNA repair activity.
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References
Collins AR (2004) The comet assay for DNA damage and repair. Mol Biotechnol 26:249–261
Shaposhnikov S, Azqueta A, Henriksson S, Meier S, Gaivão I, Brunborg G, Nilsson M, Collins AR (2010) Twelve-gel slide format optimised for comet assay and fluorescent in situ hybridisation. Tox Letters 19(195):31–34
Acknowledgment
This project was partially financed by the EU FP7 project NANoREG, grant 310584, and Research Council of Norway funded French-Norwegian project ComPack, grant 13-03 FNS.
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Shaposhnikov, S., Collins, A. (2017). Twelve-Gel Comet Assay Format for Quick Examination of DNA Damage and Repair. In: Didenko, V. (eds) Fast Detection of DNA Damage. Methods in Molecular Biology, vol 1644. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7187-9_16
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DOI: https://doi.org/10.1007/978-1-4939-7187-9_16
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7185-5
Online ISBN: 978-1-4939-7187-9
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