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© 1995 Oxford University Press

research-article

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

Vladimir J. Bykov, Rajiv Kumar, Asta Försti and Kari Hemminki

Center for Nutrition and Toxicology, Karolinska Institute Novum, 141 57 Huddinge, Sweden

UV-Induced cyclobutane dimers and 6–4 photoproducts, containing an unmodilled nucleotide at the 5'-posltion were released from DNA by means of digestion with DNase I, snake venom phosphodlesterase and prostatic acid phos phatase. The enzymes were deactivated by proteinase K followed by ethanol precipitation. The products were phos phorylated 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.


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