Could a strong alkali deproteinization replace the standard lysis step in alkaline single cell gel electrophoresis (comet) assay (pH > 13)?

Mutation Research - Genetic Toxicology and Environmental Mutagenesis

Volumn 678, Issue 1, 2009, Pages 65-70

  Vivek Kumar, P R ^a   Cheriyan, V D ^a   Seshadri, M ^a  

^a BHABHA ATOMIC RESEARCH CENTRE   (India)

Author keywords

Alkali deproteinization; Comet assay; DNA damage; Human blood lymphocytes

Indexed keywords

GENOMIC DNA;

ARTICLE; COMET ASSAY; CYTOLYSIS; DNA DAMAGE; DNA DENATURATION; DNA STRAND BREAKAGE; HUMAN; IRRADIATION; PERIPHERAL LYMPHOCYTE; PH MEASUREMENT; PRIORITY JOURNAL;

ALKALIES; COMET ASSAY; HUMANS; MALE; SODIUM HYDROXIDE;

EID: 68349158810     PISSN: 13835718     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.mrgentox.2009.06.007     Document Type: Article

References (18)

1. Tice R.R., Agurell E., Anderson D., Burlinson B., Hartmann A., Kobayashi H., Miyamae Y., Rojas E., Ryu J.-C., and Sasak Y.F. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen. 35 (2000) 206-221
2. Collins A.R. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol. Biotechnol. 26 (2004) 249-261
3. Ostling O., and Johanson K.J. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem. Biophys. Res. Commun. 123 (1984) 291-298
4. Singh N.P., McCoy M.T., Tice R.R., and Schneider E.L. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell. Res. 175 (1988) 184-191
5. Olive P.L. DNA organization affects cellular radiosensitivity and detection of initial DNA strand breaks. Int. J. Radiat. Biol. 62 (1992) 389-396
6. R. Tice, M. Vasquez, Protocol for the application of the pH > 13 alkaline single cell gel (SCG) assay to the detection of DNA damage in mammalian cells, Comet assay interest group website, http://cometassay.com/.
7. Fairbairn D.W., Olive P.L., and O'Neill K.L. The comet assay: a comprehensive review. Mutat. Res. 339 (1995) 37-59
8. Sestili P., Martinelli C., and Stocchi V. The fast halo assay: an improved method to quantify genomic DNA strand breakage at the single cell-level. Mutat. Res. 607 (2006) 205-214
9. Konca K., Lankoff A., Banasik A., Lisowska H., Kuszewski T., Gozdz S., Koza Z., and Wojcik A. A cross-platform public domain PC image-analysis program for the comet assay. Mutat. Res. 534 (2003) 15-20
10. Singh N.P., and Stephens R.E. Microgel electrophoresis: sensitivity, mechanism and DNA electrostretching. Mutat. Res. 383 (1997) 167-175
11. Rojas E., Lopez M.C., and Valverde M. Single cell gel electrophoresis assay: methodology and applications. J. Chromatogr. B 722 (1999) 225-254
12. Green M.H.L., Lowe J.E., Delaney R.A., and Green I.C. Comet assay to detect nitric oxide-dependent DNA damage in mammalian cells. Methods Enzymol. 269 (1996) 243-266
13. Cook P.R., Brazell I.A., and Jost E. Characterization of nuclear structures containing super helical DNA. J. Cell Sci. 22 (1976) 303-324
14. Collins A.R., Dobson V.l., Dusinska M., Kennedy G., and Stetina R. The comet assay: what can it really tell us?. Mutat. Res. 375 (1997) 183-193
15. Carlo D.D., Ionescu-Zanetti C., Zhang Y., Hung P., and Lee L.P. On-chip cell lysis by local hydroxide generation. Lab Chip 5 (2005) 171-178
16. Collins A.R., Dusinska M., Gedik C.M., and Stetina R. Oxidative damage to DNA: do we have a reliable biomarker?. Environ. Health Perspect. 104 (1996) 465-469
17. Collins A.R., Dusinska M., Franklin M., Somorovska M., Petrovska H., Duthie S., Fillion L., Panayiotidis M., Raslova K., and Vaughan N. Comet assay in human biomonitoring studies: reliability, validation, and application. Environ. Mol. Mutagen. 30 (1997) 139-146
18. Kumaravel T.S., and Jha A.N. Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals. Mutat. Res. 605 (2006) 7-16