This version is an update to the 24th edition of Standard Methods for the Examination of Water and Wastewater and was posted online April 18, 2025.

8071 COMET ASSAY—SINGLE-CELL GEL ELECTROPHORESIS FOR DETECTING DNA DAMAGE

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8071 A. Introduction References
1. Cooper GM. DNA repair. In: The Cell: a molecular approach. 2nd ed. Sunderland (MA): Sinauer Associates; 2000. Google Scholar
2. Sugg DW, Chesser RK, Brooks JA, Grasman BT. The association of DNA damage to concentrations of mercury and radiocesium in largemouth bass. Environ Toxicol Chem.1995;14(4):661–668. Google Scholar
3. Ralph S, Petras M, Pandrangi R, Vrzoc M. Alkaline single-cell gel (comet) assay and genotoxicity monitoring using two species of tadpoles. Environ Molecul Mutagen. 1996;28:112–120. Google Scholar
4. Sugg DW, Brooks JA, Jagoe CH, Smith MH, Chesser RK, Bickham JW, Lomakin MD, Dallas CE, Baker RJ. DNA damage and radiocesium in channel catfish from Chernobyl. Environ Toxicol Chem. 1996;15(7):1057–1063. Google Scholar
5. Mitchelmore CL, Chipman JK. DNA strand breakage in aquatic organisms and the potential value of the comet assay in environmental monitoring. Mutation Res. 1998;399(2):135–147. Google Scholar
6. Blasiak J, Jaloszynski P, Trzeciak A, Szyfter K. In vitro studies on the genotoxicity of the organophosphorus insecticide malathion and its two analogues. Mutation Res. 1999;445(2):275–283. Google Scholar
7. Park JK, Lee JS, Lee HH, Choi IS, Park SD. Accumulation of polycyclic aromatic hydrocarbon-induced single-strand breaks is attributed to slower rejoining processes by DNA polymerase inhibitor, cytosine arabinoside in CHO-K1 cells. Life Sci. 1991;48(13):1255–1261. Google Scholar
8. Eastman A, Barry MA. The origins of DNA breaks: a consequence of DNA damage, DNA repair or apoptosis? Cancer Invest. 1992;10:229–240. Google Scholar
9. Speit G, Hartmann A. The contribution of excision repair to the DNA effects seen in the alkaline single cell gel test (comet assay). Mutagenesis 1995;10(6):555–560. Google Scholar
10. Shugart L, Bickham J, Jackim G, Mcmahon G, Ridley W, Stein J, Steinert S. 1992. DNA alterations. In: Huggett RJ, Kimerle RA, Mehrle, PMJr, Bergman HL, eds. Biomarkers: Biochemical, physiological, and histological markers of anthropogenic stress. Boca Raton (FL): CRC Press; 1992. Google Scholar
11. Lee RF, Steinert SA, Nakayama K, Oshima Y. Use of DNA damage (Comet assay) and embryo development defects to assess contaminant exposure by blue crab (Callinectes sapidus) embryos. In: Henshel DS, Black MC, Harrass MC, eds. Environmental toxicology and risk assessment: Standardization of biomarkers for endocrine disruption and environmental assessment; ASTM STP 1364, p. 341. West Conshohocken (PA): ASTM International; 1999, Vol. 8. Google Scholar
12. Steinert SA. DNA damage as bivalve biomarker. Biomarkers. 1999;4(6):492–496. Google Scholar
8071 B. Comet Assay (Single-Cell Gel Electrophoresis) References
1. Lee RF, Steinert SA. Use of the single-cell electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Mutation Res. 2003;544(1):43–64. Google Scholar
2. Whitehead A, Kuivila KM, Orlando JL, Kotelevtsev S, Anderson SL. Genotoxicity in native fish associated with agriculture runoff events.Environ Toxicol Chem. 2004; 23(12):2868–2877. Google Scholar
3. Nehks S, Segner H. Comet assay with the fish cell line rainbow trout gonad-2 for in vitro genotoxicity testing for xenobotics and surface waters. Environ Toxicol Chem. 2005;24:2078–. Google Scholar
4. Coughlan BM, Hartl MGJ, O’Reilly SJ, Sheehan D, Mothersill C, van Pelt FNAM, O’Halloran J, O’Brien NM. Detecting genotoxicity using the Comet assay following chronic exposure of Manila clam Tapes semidecussatus to polluted estuarine sediments. Marine Pollut Bull. 2004;44(12):1359–1365. Google Scholar
5. Kilemade MF, Hartl MGJ, Sheehan D, Mothersill C, van Pelt FNAM, O’Halloran J, O’Brien NM. 2004. Genotoxicity of field-collected intertidal sediments from Cork Harbor, Ireland, to juvenile turbot (Scophthalmus maximus L.) as measured by the Comet assay. Environ Mutagen. 2004;44(1):56–64. Google Scholar
6. Kosmehl T, Hallare AV, Reifferscheid G, Manz W, Braunbeck T, Hollert H. 2006. A novel contact assay for testing genotoxicity of chemicals and whole sediments in zebrafish embryos. Environ Toxicol Chem. 25(8):2097–2106. Google Scholar
7. Rydberg B, Johanson KJ. Estimation of DNA strand breaks in single mammalian cells. In: Hanawalt PC, Friedberg EC, Fox CF, eds. DNA repair mechanisms. New York (NY): Academic Press; 1978, p. 465-468. Google Scholar
8. Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for the quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988;175(1):184–191. Google Scholar
9. McKelvey-Martin VJ, Green MHL, Schmezer P, Pool-Zobel BL, De Meo MP, Collins A. The single cell gel electrophoresis assay (comet assay): a European review. Mutation Res. 1993;288(1):47–63. Google Scholar
10. Pandrangi R, Petras M, Ralph S, Vrzoc M. Alkaline single cell gel (comet) assay and genotoxicity monitoring using bullheads and carp. Environ Molecul Mutagen. 1995;26(4):345–356. Google Scholar
11. Steinert SA, Streib-Montee R, Leather JM, Chadwick DB. DNA damage in mussels at sites in San Diego Bay. Mutation Res. 1988;399(1):65–85. Google Scholar
12. Steinert SA, Streib-Montee R, Sastre MP. Influence of sunlight on DNA damage in mussels exposed to polycyclic aromatic hydrocarbons. Mar Environ Res. 1988;46(1-5):355–358. Google Scholar
13. Collins AR, Duthie SJ, Dobson VL. Direct enzymic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis. 1993;14(9):1733–1735. Google Scholar
14. Tice RR. The single cell gel/comet assay: a microgel electrophoretic technique for the detection of DNA damage and repair in individual cells. In: D.H. Phillips DH, Venitt S, eds. Environmental mutagenesis. Oxford (UK): BIOS Scientific Publishers; 1995. Google Scholar
15. Villarini M, Moretti M, Damiani E, Greci L, Santroni AM, D. Fedeli D, Falcioni G. Detection of DNA damage in stressed trout nucleated erythrocytes using the comet assay: protection by nitroxide radicals. Free Rad Biol Med. 1998;24(7-8):1310–1315. Google Scholar
16. Singh NP. Microgel electrophoresis of DNA from individual cells: Principles and methodology. In: Pfeifer GP, ed. Technologies for detection of DNA damage and mutations, New York (NY): Plenum Press; 1996 p 3. Google Scholar
17. Gedik CM, Ewen SWB, Collins AR. Single-cell gel electrophoresis applied to the analysis of UV-C damage and its repair in human cells. Int J Radiat Biol. 1992;62(3):313–320. Google Scholar
18. Collins AR, Dusinska M, Gedik CM, Stetina R. Oxidative damage to DNA: do we have a reliable biomarker? Environ Health Perspect. 1996;104(suppl 3):465–469. Google Scholar
19. Leroy T. Evaluation of three methods for the detection of DNA single-strand breaks in human lymphocytes: alkaline elution, nick translation, and single-cell gel electrophoresis. J Toxicol Environ Health. 1996;47(5):409–422. Google Scholar

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Standard Methods Committee of the American Public Health Association, American Water Works Association, and Water Environment Federation. 8071 comet assay—single-cell gel electrophoresis for detecting dna damage In: Standard Methods For the Examination of Water and Wastewater. Lipps WC, Baxter TE, Braun-Howland E, editors. Washington DC: APHA Press.

DOI: 10.2105/SMWW.2882.155

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