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. 2014 Sep 9;7:9.
doi: 10.1186/s13628-014-0009-z. eCollection 2014.

Mathematical Modelling of the Automated FADU Assay for the Quantification of DNA Strand Breaks and Their Repair in Human Peripheral Mononuclear Blood Cells

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Free PMC article

Mathematical Modelling of the Automated FADU Assay for the Quantification of DNA Strand Breaks and Their Repair in Human Peripheral Mononuclear Blood Cells

Michael Junk et al. BMC Biophys. .
Free PMC article

Abstract

Background: Cells continuously undergo DNA damage from exogenous agents like irradiation or genotoxic chemicals or from endogenous radicals produced by normal cellular metabolic activities. DNA strand breaks are one of the most common genotoxic lesions and they can also arise as intermediates of DNA repair activity. Unrepaired DNA damage can lead to genomic instability, which can massively compromise the health status of organisms. Therefore it is important to measure and quantify DNA damage and its repair.

Results: We have previously published an automated method for measuring DNA strand breaks based on fluorimetric detection of alkaline DNA unwinding [1], and here we present a mathematical model of the FADU assay, which enables to an analytic expression for the relation between measured fluorescence and the number of strand breaks.

Conclusions: Assessment of the formation and also the repair of DNA strand breaks is a crucial functional parameter to investigate genotoxicity in living cells. A reliable and convenient method to quantify DNA strand breakage is therefore of significant importance for a wide variety of scientific fields, e.g. toxicology, pharmacology, epidemiology and medical sciences.

Keywords: DNA repair; DNA strand breaks; FADU; Mathematical model.

Figures

Figure 1
Figure 1
Signal fluorescence depending on irradiation dose. Human peripheral blood mononuclear cells (PBMC) were irradiated with several doses of x-ray. DNA strand breaks formation was measured using the automated FADU assay exactly as described [1].The Sybr-green fluorescence intensity, which is a direct marker of double-stranded DNA, decreases with increasing the irradiation dose, due to progressive alkaline unwinding of the DNA in the lysate starting from DNA ends or breaks. Circles represent the mean of three experimental replicates. Each graph represents the one donor.

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