Differences in detection of DNA adducts in the 32P-postlabelling assay after either 1-butanol extraction or nuclease P1 treatment.
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.
Gallagher
J E
JE
Genetic Toxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711.
Jackson
M A
MA
George
M H
MH
Lewtas
J
J
Robertson
I G
IG
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
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