Effect of tissue fixatives on telomere length determination by quantitative PCR.
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.
Koppelstaetter
Christian
C
Division of Nephrology, Department of Internal Medicine, Medical University Innsbruck, Anichstrasse 35, Innsbruck 6020, Austria. christian.koppelstaetter@uibk.ac.at
Jennings
Paul
P
Hochegger
Kathrin
K
Perco
Paul
P
Ischia
Rudolf
R
Karkoszka
Henryk
H
Mayer
Gert
G
D016922:Cellular Senescence; D017931:DNA Primers / Q000737:chemistry; D005404:Fixatives / Q000494:pharmacology*; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D010232:Paraffin / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016615:Telomere / Q000187:drug effects / Q000648:ultrastructure*; D013997:Time Factors
D017931:DNA Primers; D005404:Fixatives; D005557:Formaldehyde; D012313:RNA; D010232:Paraffin