Different qPCR master mixes influence telomere primer binding within and between bird species

The analysis of telomere dynamics in birds is a growing research field providing important findings on ecological and environmental variations in individuals' aging, fitness and lifespan. Real‐time quantitative PCR (qPCR) has gained much interest for the evaluation of telomere length in birds. However, the assessment of several key preanalytical and analytical factors to optimize the method for achieving reproducible results, and the influence of these factors on the conclusions of each study, have been generally overlooked. In this study, we assessed the performance of eight commercially‐available qPCR master mixes in the amplification of telomere fragments in two bird species (zebra finch Taeniopygia guttata, and red‐billed chough Pyrrhocorax pyrrhocorax). We observed that qPCR master mixes influence the telomere primer binding to target sequences and the amplification specificity within and between bird species, although PCR amplification efficiencies were very close to 100% for all ...