Nonradiometric detection of DNA crosslinks in mussel hemolymph by alkaline elution.
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.
Batel
R
R
Ruder Bosković Institute, Center for Marine Research, Rovinj, Croatia.
Vukmirović
M
M
Bihari
N
N
Zahn
R K
RK
Müller
W E
WE
D000818:Animals; D049872:Bivalvia / Q000737:chemistry* / Q000187:drug effects; D001761:Bleomycin / Q000378:metabolism / Q000633:toxicity; D002623:Chemistry Techniques, Analytical / Q000379:methods; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D004249:DNA Damage; D005296:Ferrous Compounds / Q000378:metabolism / Q000633:toxicity; D005557:Formaldehyde / Q000633:toxicity; D006458:Hemolymph / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D016685:Mitomycin / Q000633:toxicity; D011874:Radiometry; D015203:Reproducibility of Results
D005296:Ferrous Compounds; D001761:Bleomycin; D005557:Formaldehyde; D016685:Mitomycin; D004247:DNA