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Results from the “Technical Workshop on Genotoxicity Biosensing” on the micro-scale fluorometric assay of deoxyribonucleic acid unwinding

Baumstark-Khan, Christa and Horneck, Gerda (2007) Results from the “Technical Workshop on Genotoxicity Biosensing” on the micro-scale fluorometric assay of deoxyribonucleic acid unwinding. Analytica Chimica Acta, 593, pp. 75-81. Elsevier.

Full text not available from this repository.

Abstract

The fluorometric analysis of DNA unwinding (FADU assay) was originally designed for rapid detection of X-ray-induced DNA damage in mammalian cells. This cellular bioassay is based on time-dependent alkaline denaturation of DNA under moderate denaturing conditions (pH 12.2–12.4) starting from ends as well as from all DNA break points (single-strand breaks, SSB; double-strand breaks, DSB; alkali-labile sites, ALS). DNA which remained double-stranded after 30 min of alkaline treatment was detected after neutralisation and immediate fragmentation followed by binding to the Hoechst 33258 dye (bisbenzimide) and fluorescence measures. In the current paper, a modified method was used which allows cell cultivation and chemical treatment in the same microplate (μ-FADU) followed by analysis of 96 samples in a microplate fluorescence reader. Exposure of mammalian cells to chemicals was performed for 60 min on ice thus allowing identification of direct acting substances capable of inducing DNA-strand breaks. As an inter-assay standard the action of hydrogen peroxide was tested in every test plate. The results demonstrate that the μ-FADU assay is suitable to detect the presence of chemically induced strand breaks within 3 h.

Item URL in elib:https://elib.dlr.de/51563/
Document Type:Article
Title:Results from the “Technical Workshop on Genotoxicity Biosensing” on the micro-scale fluorometric assay of deoxyribonucleic acid unwinding
Authors:
AuthorsInstitution or Email of AuthorsAuthors ORCID iD
Baumstark-Khan, ChristaUNSPECIFIEDUNSPECIFIED
Horneck, GerdaUNSPECIFIEDUNSPECIFIED
Date:2007
Journal or Publication Title:Analytica Chimica Acta
Refereed publication:Yes
Open Access:No
Gold Open Access:No
In SCOPUS:No
In ISI Web of Science:Yes
Volume:593
Page Range:pp. 75-81
Publisher:Elsevier
Status:Published
Keywords:Chinese hamster ovary cells, Fluorometric analysis of deoxyribonucleic acid unwinding, Chemical exposure, Deoxyribonucleic acid strand breaks, Technical Workshop on Genotoxicity Biosensing workshop
HGF - Research field:Aeronautics, Space and Transport (old)
HGF - Program:Space (old)
HGF - Program Themes:W FR - Forschung unter Weltraumbedingungen (old)
DLR - Research area:Space
DLR - Program:W FR - Forschung unter Weltraumbedingungen
DLR - Research theme (Project):W - Vorhaben Strahlenbiologie (old)
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Radiation Biology
Deposited By: Kopp, Kerstin
Deposited On:12 Oct 2007
Last Modified:27 Apr 2009 14:25

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