32P-Postlabelling: A Sensitive Technique for the Detection of DNA Adducts
aDepartment of Biochemistry, Faculty of Science, Charles University, Prague, bDepartment of Molecular Toxicology, German Cancer Research Centre, Heidelberg, Germany
The 32P-postlabelling assay has emerged as a major method to detect DNA adducts induced by structurally diverse carcinogens and other DNA lesions. The technique comprises enzymatic degradation of DNA to 3'-monodeoxynucleotides, enrichment of adducts, 5'-32P-labelling, adduct separation by TLC (or HPLC), and detection and quantitation of adducts. The review concerns individual 32P-postlabelling techniques (standard procedure, enrichment methods) as well as the applications of the method to different DNA modifications. Recent reports on the application of the 32P-postlabelling technique in human risk assessment using carcinogen-DNA adducts as molecular biomarkers are discussed and a critical evaluation of the assays is presented.
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