title: Some quantitative considerations about DNA adduct enrichment procedures for 32P-postlabelling.
abstract: The concentrations of 2'-deoxyribonucleoside-3'-monophosphates remaining in calf-thymus DNA digests after nuclease P1 digestion or extraction into 1-butanol, the most commonly used adduct enrichment procedures prior to the application of the 32P-postlabelling assay, were measured using HPLC and 32P-postlabelling methods. When 10 micrograms of DNA digested to mononucleotides was used, the total amount of nucleotides remaining in the samples were approximately 4 and approximately 14 pmol after nuclease P1 treatment or 1-butanol extraction respectively. The influence of various concentrations of normal nucleotides on the labelling efficiency of a 2'-deoxyguanosine-3'-monophosphate adduct of benzo[a]pyrene diol-epoxide was also studied and found to depend upon the ratio of normal nucleotides/adducted nucleotides present in the sample. Also, the ATP/normal nucleotides ratio in the phosphorylation reaction may affect the quantitation of the adducts and thus deserves due consideration.
authors: Molecular Epidemiology Unit, Center for Nutrition and Toxicology, NOVUM, Karolinska Institute, Huddinge, Sweden.
authors: C E
authors: CE
authors: Vaca
authors: M
authors: M
authors: Löfgren
authors: K
authors: K
authors: Hemminki
mesh_terms: D020001:1-Butanol; D000818:Animals; D001564:Benzo(a)pyrene / Q000737:chemistry; D000440:Butanols; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D018736:DNA Adducts; D004249:DNA Damage; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D009711:Nucleotides / Q000032:analysis / Q000737:chemistry; D010761:Phosphorus Radioisotopes; D015719:Single-Strand Specific DNA and RNA Endonucleases
chemical_list: D000440:Butanols; D018736:DNA Adducts; D003838:Deoxyadenine Nucleotides; D009711:Nucleotides; D010761:Phosphorus Radioisotopes; C056612:benzo(a)pyrene-DNA adduct; D001564:Benzo(a)pyrene; C012873:2'-deoxy-5'-adenosine monophosphate; D020001:1-Butanol; D004247:DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases