title: Differences in detection of DNA adducts in the 32P-postlabelling assay after either 1-butanol extraction or nuclease P1 treatment.
abstract: The use of nuclease P1 treatment and 1-butanol extraction to increase the sensitivity of the 32P-postlabelling assay for DNA adducts have been compared. Although similar results were obtained with the two methods for standard adducts formed with benzo[a]pyrene diol epoxide I (BPDE-I), nuclease P1 treatment resulted in a significant reduction in detection of major adducts from 1-amino-6-nitropyrene (1-amino-6-NP), 1-amino-8-nitropyrene (1-amino-8-NP), 2-aminofluorene (2-AF), 2-naphthylamine (2-NA) and 4-aminobiphenyl (4-ABP) modified DNAs, but not following the 32P-postlabelling analysis of 2-acetylaminofluorene (2-AAF) modified DNA. These results suggest that, at least initially, both modifications of the 32P-postlabelling assay should be used for the detection of unknown adducts or for adducts derived from nitroaromatics and aromatic amines.
authors.affiliation: Genetic Toxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711.
authors.forename: J E
authors.initials: JE
authors.lastname: Gallagher
authors.forename: M A
authors.initials: MA
authors.lastname: Jackson
authors.forename: M H
authors.initials: MH
authors.lastname: George
authors.forename: J
authors.initials: J
authors.lastname: Lewtas
authors.forename: I G
authors.initials: IG
authors.lastname: Robertson
mesh_terms: D020001:1-Butanol; D000255:Adenosine Triphosphate / Q000378:metabolism; D001345:Autoradiography / Q000379:methods; D000440:Butanols; D002273:Carcinogens / Q000032:analysis / Q000378:metabolism*; D002855:Chromatography, Thin Layer / Q000379:methods; D004247:DNA / Q000032:analysis / Q000378:metabolism*; D009708:Nucleotidases; D010761:Phosphorus Radioisotopes; D015719:Single-Strand Specific DNA and RNA Endonucleases
chemical_list: D000440:Butanols; D002273:Carcinogens; D010761:Phosphorus Radioisotopes; D000255:Adenosine Triphosphate; D020001:1-Butanol; D004247:DNA; D009708:Nucleotidases; D015719:Single-Strand Specific DNA and RNA Endonucleases