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Int. J. Mol. Sci. 2013, 14(6), 11544-11559; doi:10.3390/ijms140611544
Communication

Apparent Polyploidization after Gamma Irradiation: Pitfalls in the Use of Quantitative Polymerase Chain Reaction (qPCR) for the Estimation of Mitochondrial and Nuclear DNA Gene Copy Numbers

1,2,* , 1
, 1
, 1,3
 and 1,2,4
1 Australian Nuclear Science and Technology Organisation, Lucas Heights, Sydney, New South Wales 2234, Australia 2 Medical Radiation Sciences, Faculty of Health Sciences, University of Sydney, Cumberland, Sydney, New South Wales 2141, Australia 3 School of Physics, University of Sydney, Camperdown, Sydney, New South Wales 2006, Australia 4 National Imaging Facility at Brain and Mind Research Institute (BMRI), University of Sydney, Camperdown, Sydney, New South Wales 2050, Australia
* Author to whom correspondence should be addressed.
Received: 5 March 2013 / Revised: 18 April 2013 / Accepted: 16 May 2013 / Published: 30 May 2013
(This article belongs to the Section Molecular Pathology)
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Abstract

Quantitative polymerase chain reaction (qPCR) has been widely used to quantify changes in gene copy numbers after radiation exposure. Here, we show that gamma irradiation ranging from 10 to 100 Gy of cells and cell-free DNA samples significantly affects the measured qPCR yield, due to radiation-induced fragmentation of the DNA template and, therefore, introduces errors into the estimation of gene copy numbers. The radiation-induced DNA fragmentation and, thus, measured qPCR yield varies with temperature not only in living cells, but also in isolated DNA irradiated under cell-free conditions. In summary, the variability in measured qPCR yield from irradiated samples introduces a significant error into the estimation of both mitochondrial and nuclear gene copy numbers and may give spurious evidence for polyploidization.
Keywords: gamma radiation; temperature; mitochondria; nucleus; DNA copy number; polyploidization gamma radiation; temperature; mitochondria; nucleus; DNA copy number; polyploidization
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Kam, W.W.Y.; Lake, V.; Banos, C.; Davies, J.; Banati, R. Apparent Polyploidization after Gamma Irradiation: Pitfalls in the Use of Quantitative Polymerase Chain Reaction (qPCR) for the Estimation of Mitochondrial and Nuclear DNA Gene Copy Numbers. Int. J. Mol. Sci. 2013, 14, 11544-11559.

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