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RECORD NUMBER: 14 OF 586

Main Title 32P-Adduct Assay: Comparative Recoveries of Structurally Diverse DNA Adducts in the Various Enhancement Procedures.
Author Gupta, R. C. ; Earley, K. ;
CORP Author Baylor Coll. of Medicine, Houston, TX. Dept. of Pharmacology.;Health Effects Research Lab., Research Triangle Park, NC.
Publisher c1988
Year Published 1988
Report Number EPA/600/J-88/466;
Stock Number PB90-147711
Additional Subjects Carcinogens ; Deoxyribonucleic acids ; Phosphorus isotopes ; Humans ; Thin layer chromatography ; Nucleosides ; Nucleotides ; Reprints ; DNA damage ; Deoxyribonucleases ; Polynucleotide hydroxylkinase
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Status
NTIS  PB90-147711 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 9p
Abstract
A (32)P-adduct assay for the measurement of low levels (1 adduct per 10(sup 7) nucleotides) of binding of carcinogens to DNA has been reported previously. In this procedure, DNA is enzymatically hydrolyzed to 3'-monophosphates of normal nucleosides and adducts, which are 5'-(32)P-labeled by T4 polynucleotide kinase and (lambda(32)P)ATP. Labeled adducts are resolved by TLC. Enrichment of adducts by extraction in 1-butanol or digestion with nuclease P1 prior to (32)P-labeling, however, increased the sensitivity of detection for many adducts to a level of 1 per 10(sup 9-10) nucleotides, although adduct recovery particularly in the latter assay depended on the chemical nature of adducts. The observation that chemical structure of an adduct may be detrimental in its recovery in the enzyme- and extraction-mediated enrichment procedures may serve as a probe in the structural characterization of adducts of unknown carcinogens. (Copyright 1988 IRL Press Ltd., Oxford, England.)