Title
Improvements in the 32P-postlabelling procedure to quantify bulky aromatic DNA adducts
Author
Steenwinkel, M.J.S.T.
Roggeband, R.
van Delft, J.H.M.
Baan, R.A.
Contributor
Phillips, D.H. (editor)
Castegnaro, M. (editor)
Bartsch, H. (editor)
Publication year
1993
Abstract
This contribution describes methodological modifications and improvements that may contribute to inter-assay reproducibility and more accurate adduct quantification for 32P-postlabelling. Firstly, an anion-exchange chromatography procedure was developed to determine the amount of DNA used per assay and to check its purity, in particular to verify the absence of contaminating RNA. Secondly, calibration standards were prepared, in order to correct for differences in recovery. The modification levels of these standards were determined by synchronous fluorescence spectrophotometric analysis. Thirdly, the effect on adduct levels of exposure to light during postlabelling was investigated. Exposure of polyaromatic DNA adducts on a PEI-cellulose plate reduced the amounts of adducts detected considerably. Chemicals/CAS: DNA, 9007-49-2; phosphorus, 7723-14-0; RNA, 63231-63-0; Carcinogens; DNA, 9007-49-2; Phosphorus Radioisotopes; Polycyclic Hydrocarbons; RNA, 63231-63-0
Subject
Phosphorus
Polycyclic hydrocarbon
Ion exchange chromatography
Isolation and purification
Light
Methodology
Reproducibility
Standard
Carcinogens
Chromatography, Ion Exchange
DNA
DNA Damage
Environmental Monitoring
Evaluation Studies
Human
Light
Occupational Exposure
Phosphorus Radioisotopes
Polycyclic Hydrocarbons
Reference Standards
Reproducibility of Results
RNA
To reference this document use:
http://resolver.tudelft.nl/uuid:7a065a2f-73f7-4ae3-8a19-e18b0e8966ea
TNO identifier
82307
Publisher
IARC, Lyon
Source
Postlabelling methods for detection of DNA adducts, 65-70
Series
IARC Scientific Publications
Bibliographical note
Human and Experimental Toxicology, (12)4, 351
Document type
bookPart