A quantitative real-time PCR method for absolute telomere length.
Telomere shortening is an important risk factor for cancer and accelerated aging. Here we describe the development of a simple and reproducible method to measure absolute telomere length. Based on Cawthon's quantitative real-time PCR (qRT-PCR) assay, our method uses an oligomer standard that can be used to generate absolute telomere length values rather than relative quantification. We demonstrate a strong correlation between this improved method and the "gold standard" of telomere length measurement-terminal restriction fragment analysis (TRF) by Southern hybridization. The capability to generate absolute telomere length values should allow a more direct comparison of results between experiments within and between laboratories.
O'Callaghan
Nathan
N
Commonwealth Scientific and Industrial Research Organization (CSIRO)-Human Nutrition, Adelaide, Australia. nathan.o'callaghan@csiro.au
Dhillon
Varinderpal
V
Thomas
Philip
P
Fenech
Michael
M
D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D016615:Telomere / Q000235:genetics* / Q000648:ultrastructure*