Real-time quantitative PCR of telomere length

Telomeres cap the ends of chromosomes and are essential for the protection of chromosomes, as well as restricting the replicative potential, of a cell These functions are achieved by the regulation of repeat length, making the measurement of telomere length a useful, aid in the elucidation of the replicative history and potential of cells. Previously published techniques-employed either hybridization or flow cytometry methods, which Are technically demanding and time-consuming. In 2002, R. M. Cawthon published a real-time polymerase chain reaction (PCR)-based,method for telolmere length measurement using the Applied Biosystems Prism, 7700,sequence detection system. The technique measures the factor by which the ratio of telomere repeats copy number to single-gene copy number differs between a sample and that of a reference deoxyribonucleic acid sample. In many laboratories worldwide, including ours, real-time PCR is carried out using the Roche LightCycler; as opposed to the AB Prism system. This benchmark details the modifications to Cawthon's method, and describes the parameters and reagents required to measure telomere length using the Roche LightCycler.