Abstract
Comet assay provides a rapid and reliable way to screen genotoxic effects of a wide variety of nanoparticles. Single cell gel electrophoresis assay (SCGE) or comet assay gained in popularity as a standard technique and is widely used for testing novel particles or chemicals for genotoxicity, monitoring environmental contamination with genotoxins, human biomonitoring and molecular epidemiology, and fundamental research in DNA damage and repair. Here, we describe the method of alkaline comet assay to detect nanoparticle-induced DNA damage.
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References
Geraci CL, Castranova V (2010) Challenges in assessing nanomaterial toxicology: a personal perspective. Wiley Interdiscip Rev Nanomed Nanobiotechnol 2:569–577
Kisin ER, Murray AR, Keane MJ, Shi XC, Schwegler-Berry D, Gorelik O, Arepalli S, Castranova V, Wallace WE, Kagan VE, Shvedova AA (2007) Single-walled carbon nanotubes: geno- and cytotoxic effects in lung fibroblast V79 cells. J Toxicol Environ Health A 70:2071–2079
Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191
Fairbairn DW, Olive PL, O’Neill KL (1995) The comet assay: a comprehensive review. Mutation Res 339:37–59
Lemay M, Wood KA (1999) Detection of DNA damage and identification of UV-induced photoproducts using the Comet Assay® kit. Biotechniques 27(4):846–851
Malyapa RS, Bi C, Ahern EW, Roti JL (1998) Detection of DNA damage by the alkali comet assay after exposure to low dose gamma radiation. Radiat Res 149:396–400
Henderson L, Wolfreys A, Fedyk J, Bourner C, Windeback S (1998) The ability for the comet assay to discriminate between genotoxins and cytotoxins. Mutagenesis 13:89–94
Visvardis EE, Tassiou AM, Piperakis SM (1997) Study of DNA damage induction and repair capacity of fresh cryopreserved lymphocytes exposed to H2O2 and γ-irradiation with the alkaline comet assay. Mutat Res 383:71–80
Collins AR, Ma AG, Duthie SJ (1995) The kinetics of repair of oxidative DNA damage (strand breaks and oxidized pyrimidine dimers) in human cells. Mutation Res 336:69–77
Singh NP, Stephens RE (1997) Microgel electrophoresis: sensitivity, mechanisms, and DNA electrostretching. Mutation Res 383:167–175
Östling O, Johanson KJ (1984) Microelectrophoretic study of radiation-induced DNA damage in individual cells. Biochem Biophys Res Commun 123:291–298
Morris EJ, Dreixler JC, Cheng K-Y, Wilson PM, Gin RM, Geller HM (1999) Optimization of single-cell gel electrophoresis (SCGE) for quantitative analysis of neuronal DNA damage. Biotechniques 26:282–289
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Bowman, L., Castranova, V., Ding, M. (2012). Single Cell Gel Electrophoresis Assay (Comet Assay) for Evaluating Nanoparticles-Induced DNA Damage in Cells. In: Soloviev, M. (eds) Nanoparticles in Biology and Medicine. Methods in Molecular Biology, vol 906. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-953-2_34
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DOI: https://doi.org/10.1007/978-1-61779-953-2_34
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Publisher Name: Humana Press, Totowa, NJ
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