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Differences in detection of DNA adducts in the sup 32 P-postlabelling assay after either 1-butanol extraction or nuclease Pl treatment

Technical Report ·
OSTI ID:6140928
The use of nuclease P1 treatment and 1-butanol extraction to increase the sensitivity of the {sup 32}P-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, nuclease P1 treatment resulted in a significant reduction in detection of major adducts 1-amino-6-nitropyrene, 1-amino-8-nitropyrene, 2-aminofluorene, 2-naphthylamine and 4-aminobiphenyl modified DNAs, but not following the {sup 32}P-postlabelling analysis of 2-acetylaminofluorene modified DNA. These results suggest that at least initially, both modications of the {sup 32}P-postlabelling assay should be used for the detection of unknown adducts or for adducts derived from nitro-aromatics and aromatic amines.
Research Organization:
Environmental Protection Agency, Research Triangle Park, NC (USA). Health Effects Research Lab.
OSTI ID:
6140928
Report Number(s):
PB-91-115972/XAB; EPA--600/J-89/471
Country of Publication:
United States
Language:
English