title: 32P-postlabelling analysis of bulky aromatic adducts.
abstract: The 32P-postlabelling assay has emerged as a major tool for detecting DNA adducts induced by structurally diverse carcinogens, particularly bulky aromatics. The assay comprises enzymatic degradation of DNA to 3'-mononucleotides, enrichment of adducts, 5'-32P-labelling, adduct separation by TLC, and detection and quantitation of adducts. This report describes (1) an improved solvent extraction procedure for isolating DNA from tissues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios issues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios.
authors.affiliation: Preventive Medicine and Environmental Health, University of Kentucky, Lexington 40506.
authors.forename: R C
authors.initials: RC
authors.lastname: Gupta
mesh_terms: D002274:Carcinogens, Environmental / Q000032:analysis* / Q000737:chemistry; D002855:Chromatography, Thin Layer; D004247:DNA / Q000032:analysis*; D004249:DNA Damage; D006801:Humans; D008722:Methods; D010761:Phosphorus Radioisotopes; D012997:Solvents
chemical_list: D002274:Carcinogens, Environmental; D010761:Phosphorus Radioisotopes; D012997:Solvents; D004247:DNA