Effect of master mixes on the measurement of telomere length by qPCR

Abstract

Alterations in telomere length (TL) have been associated with several diseases and a method based on qPCR, the Monochrome Multiplex Real-Time Quantitative PCR (MMQPCR) technique, has been used extensively for the analysis of TL. Some previous studies have been found that certain methodological conditions can affect the measurement of TL. The aim of the study was to evaluate the performance of eight different commercially available SYBR Green and High-Resolution Melting (HRM) mixes on the measurement of TL by the MMQPCR method. Four SYBR Green and four HRM mixes were tested and the measurement of TL was expressed by the T/S ratio. It was found that the type of master mix used in MMQPCR influences the measurement of TL, affecting aspects such as the specificity and consistency of the results. Our results are the first description of the effects of different master mixes on TL analysis by MMQPCR and highlight the importance of the future methodological improvement of this broadly used technique.