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 ...鸟类端粒动力学分析是一门快速发展的研究领域，其在个体衰老、适合度与寿命的生态及环境差异研究中取得了诸多重要成果。实时定量聚合酶链式反应（real-time quantitative PCR，qPCR）已被广泛应用于鸟类端粒长度的检测。然而，当前研究普遍忽视了对若干关键分析前与分析阶段因素的评估——这些因素是优化实验方法以获得可重复结果的核心，同时也会对各项研究的结论产生显著影响。本研究针对两种鸟类（斑胸草雀*Taeniopygia guttata*与红嘴山鸦*Pyrrhocorax pyrrhocorax*），评估了8种市售qPCR预混液在端粒片段扩增中的表现。研究发现，尽管所有实验组的PCR扩增效率均接近100%，但qPCR预混液会影响端粒引物与靶序列的结合效果，以及不同鸟类物种内部及物种间的扩增特异性，虽然所有...