Differences in detection of DNA adducts in the 32P-postlabelling assay after either 1-butanol extraction or nuclease P1 treatment

Cancer Lett. 1989 Apr;45(1):7-12. doi: 10.1016/0304-3835(89)90029-3.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 1-Butanol
  • Adenosine Triphosphate / metabolism
  • Autoradiography / methods
  • Butanols
  • Carcinogens / analysis
  • Carcinogens / metabolism*
  • Chromatography, Thin Layer / methods
  • DNA / analysis
  • DNA / metabolism*
  • Nucleotidases
  • Phosphorus Radioisotopes
  • Single-Strand Specific DNA and RNA Endonucleases

Substances

  • Butanols
  • Carcinogens
  • Phosphorus Radioisotopes
  • Adenosine Triphosphate
  • 1-Butanol
  • DNA
  • Nucleotidases
  • Single-Strand Specific DNA and RNA Endonucleases