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Fast Detection of DNA Damage pp 75–93Cite as

The Fast-Halo Assay for the Detection of DNA Damage

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1644))

Abstract

The need for express screening of the DNA damaging potential of chemicals has progressively increased over the past 20 years due to the wide number of new synthetic molecules to be evaluated, as well as the adoption of more stringent chemical regulations such as the EU REACH and risk reduction politics. In this regard, DNA diffusion assays such as the microelectrophoretic comet assay paved the way for rapid genotoxicity testing. A more significant simplification and speeding up of the experimental processes was achieved with the fast halo assay (FHA) described in the present chapter. FHA operates at the single cell level and relies on radial dispersion of the fragments of damaged DNA from intact nuclear DNA. The fragmented DNA is separated by diffusion in an alkaline solvent and is stained, visualized, and finally quantified using computer-assisted image analysis programs. This permits the rapid assessment of the extent of DNA breakage caused by different types of DNA lesions. FHA has proven to be sensitive, reliable, and flexible. This is currently one of the simplest, cheapest, and quickest assays for studying DNA damage and repair in living cells. It does not need expensive reagents or electrophoretic equipment and requires only 40 min to prepare samples for computer-based quantification. This technique can be particularly useful in rapid genotoxicity assessments and in high-throughput genotoxicity screenings.

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Authors and Affiliations

  1. Dipartimento di Scienze Biomolecolari, Università degli Studi di Urbino “Carlo Bo”, Via Saffi, 2, 61029, Urbino, Italy

    Piero Sestili, Cinzia Calcabrini, Anna Rita Diaz & Vilberto Stocchi

  2. Dipartimento di Scienze per la Qualità della Vita, Alma Mater Studiorum - Università di Bologna, Corso D’Augusto 237, 47921, Rimini, Italy

    Cinzia Calcabrini & Carmela Fimognari

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  1. Piero Sestili
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  2. Cinzia Calcabrini
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  3. Anna Rita Diaz
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  4. Carmela Fimognari
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  5. Vilberto Stocchi
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Correspondence to Piero Sestili .

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Editors and Affiliations

  1. Department of Neurosurgery and Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas, USA

    Vladimir V. Didenko

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Sestili, P., Calcabrini, C., Diaz, A.R., Fimognari, C., Stocchi, V. (2017). The Fast-Halo Assay for the Detection of DNA Damage. 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_6

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  • DOI: https://doi.org/10.1007/978-1-4939-7187-9_6

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