Analysis of UV-induced DNA photoproducts by 32P-postlabelling

Carcinogenesis. 1995 Jan;16(1):113-8. doi: 10.1093/carcin/16.1.113.

Abstract

UV-induced cyclobutane dimers and 6-4 photoproducts, containing an unmodified nucleotide at the 5'-position were released from DNA by means of digestion with DNase I, snake venom phosphodiesterase and prostatic acid phosphatase. The enzymes were deactivated by proteinase K followed by ethanol precipitation. The products were phosphorylated by polynucleotide kinase and [gamma-32P]ATP. The TLC system used for the analysis enables separation of the different photoproducts and detection at a fmol level. T4 endonuclease treatment was applied to confirm the positions of cyclobutane dimers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid
  • Chromatography, Thin Layer
  • DNA / analysis
  • DNA / metabolism
  • DNA / radiation effects*
  • DNA Adducts / analysis*
  • DNA Adducts / isolation & purification
  • DNA Adducts / radiation effects
  • Humans
  • Isotope Labeling
  • Phosphorus Radioisotopes
  • Photochemistry
  • Pyrimidine Dimers / analysis
  • Ultraviolet Rays

Substances

  • DNA Adducts
  • Phosphorus Radioisotopes
  • Pyrimidine Dimers
  • DNA