Nonradiometric detection of DNA crosslinks in mussel hemolymph by alkaline elution

Anal Biochem. 1993 Aug 1;212(2):402-6. doi: 10.1006/abio.1993.1347.

Abstract

The use of bleomycin-Fe(II) complex in the determination of DNA crosslinks by alkaline elution was investigated for the purpose of developing a simple nonradiometric method applicable for detection of DNA damage in marine organisms. The assay involves preparation of active bleomycin-Fe(II) complex and its application in the washing step of the original alkaline elution procedure. Treatment of cell-free DNA from Friend leukemia cells and mussel hemolymph with bleomycin-Fe(II) complex directly on the filter resulted in a concentration-dependent strand scission response with excellent correlation (r = 0.94 and 0.98, respectively). The procedure was validated by determination of DNA cross-links in mussels injected with mitomycine C and formaldehyde.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bivalvia / chemistry*
  • Bivalvia / drug effects
  • Bleomycin / metabolism
  • Bleomycin / toxicity
  • Chemistry Techniques, Analytical / methods
  • DNA / analysis*
  • DNA / metabolism
  • DNA Damage*
  • Ferrous Compounds / metabolism
  • Ferrous Compounds / toxicity
  • Formaldehyde / toxicity
  • Hemolymph / chemistry*
  • Hemolymph / metabolism
  • Hydrogen-Ion Concentration
  • Mitomycin / toxicity
  • Radiometry
  • Reproducibility of Results

Substances

  • Ferrous Compounds
  • Bleomycin
  • Formaldehyde
  • Mitomycin
  • DNA