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

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dc.contributor.authorKam, WWYen_AU
dc.contributor.authorLake, Ven_AU
dc.contributor.authorBanos, Cen_AU
dc.contributor.authorDavies, JBen_AU
dc.contributor.authorBanati, RBen_AU
dc.date.accessioned2013-10-29T01:13:01Zen_AU
dc.date.available2013-10-29T01:13:01Zen_AU
dc.date.issued2013-06-01en_AU
dc.date.statistics2013-10-29en_AU
dc.description.abstractQuantitative 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.© 2013, MDPI Publishingen_AU
dc.identifier.citationKam, W. W. Y., Lake, V., Banos, C., Davies, J., & Banati, R. (2013). 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. International Journal of Molecular Sciences, 14 (6), 11544-11559. doi:10.3390/ijms140611544en_AU
dc.identifier.govdoc5068en_AU
dc.identifier.issn1422-0067en_AU
dc.identifier.issue6en_AU
dc.identifier.journaltitleInternational Journal of Molecular Sciencesen_AU
dc.identifier.pagination11544-11559en_AU
dc.identifier.urihttp://dx.doi.org/10.3390/ijms140611544en_AU
dc.identifier.urihttp://apo.ansto.gov.au/dspace/handle/10238/4863en_AU
dc.identifier.volume14en_AU
dc.language.isoenen_AU
dc.publisherMDPI AGen_AU
dc.subjectGamma radiationen_AU
dc.subjectMitochondriaen_AU
dc.subjectDNAen_AU
dc.subjectFragmentationen_AU
dc.subjectPolymerase chain reactionen_AU
dc.subjectPolyploidyen_AU
dc.titleApparent polyploidization after gamma irradiation: pitfalls in the use of quantitative polymerase chain reaction (qPCR) for the estimation of mitochondrial and nuclear DNA gene copy numbersen_AU
dc.typeJournal Articleen_AU
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