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Effect of tissue fixatives on telomere length determination by quantitative PCR
Authors:Koppelstaetter Christian  Jennings Paul  Hochegger Kathrin  Perco Paul  Ischia Rudolf  Karkoszka Henryk  Mayer Gert
Institution:Division of Nephrology, Department of Internal Medicine, Medical University Innsbruck, Anichstrasse 35, Innsbruck 6020, Austria. christian.koppelstaetter@uibk.ac.at
Abstract:Telomere length is a well established marker of cellular senescence and thus biological age. Quantitative PCR allows the determination even from very low amounts of tissue by using telomere specific and single copy gene primers. Comparing a directly processed tissue sample to a 4% formaldehyde fixed one showed a significantly reduced efficiency of PCR reactions (mainly in single copy gene experiments) in a storage time-dependent manner resulting in an artificial increase in reported relative telomere length. This effect was not seen when the tissue was stored in RNA later solution. In summary, telomere length determination from formaldehyde fixed material by quantitative PCR is not a reliable method. Unfortunately therefore, many easily accessible tissue samples from pathology laboratories are unsuitable for this technique.
Keywords:Formaldehyde  RNA later  Quantitative PCR  Real time PCR  Telomere length determination
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