Telomere length dynamics are an established biomarker of health and ageing in animals. The study of telomeres in numerous species has been facilitated by methods to measure telomere length by real‐time quantitative PCR (qPCR). In this method, telomere length is determined by quantifying the amount of telomeric DNA repeats in a sample and normalizing this to the total amount of genomic DNA. This normalization requires the development of genomic reference primers suitable for qPCR, which remains challenging in nonmodel organism with genomes that have not been sequenced. Here we report reference primers that can be used in qPCR to measure telomere lengths in any vertebrate species. We designed primer pairs to amplify genetic elements that are highly conserved between evolutionarily distant taxa and tested them in species that span the vertebrate tree of life. We report five primer pairs that meet the specificity and reproducibility standards of qPCR. In addition, we demonstrate an approach to choose the best primers for a given species by testing the primers on multiple individuals within a species and then applying an established computational tool. These reference primers can facilitate qPCR‐based telomere length measurements in any vertebrate species of ecological or economic interest.

中文翻译：

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针对脊椎动物中基于 qPCR 的端粒长度测量而优化的高度保守元件的引物。

端粒长度动态是动物健康和衰老的既定生物标志物。通过实时定量 PCR (qPCR) 测量端粒长度的方法促进了对许多物种端粒的研究。在这种方法中，端粒长度是通过量化样本中端粒 DNA 重复的数量并将其标准化为基因组 DNA 的总量来确定的。这种标准化需要开发适用于 qPCR 的基因组参考引物，这在基因组尚未测序的非模式生物中仍然具有挑战性。在这里，我们报告了可用于 qPCR 以测量任何脊椎动物物种的端粒长度的参考引物。我们设计了引物对来扩增在进化上相距遥远的分类群之间高度保守的遗传元件，并在跨越脊椎动物生命树的物种中进行了测试。我们报告了满足 qPCR 特异性和重现性标准的五个引物对。此外，我们通过在一个物种内的多个个体上测试引物，然后应用已建立的计算工具，展示了一种为给定物种选择最佳引物的方法。这些参考引物可以促进任何具有生态或经济利益的脊椎动物物种中基于 qPCR 的端粒长度测量。我们通过在一个物种内的多个个体上测试引物，然后应用已建立的计算工具，展示了一种为给定物种选择最佳引物的方法。这些参考引物可以促进任何具有生态或经济利益的脊椎动物物种中基于 qPCR 的端粒长度测量。我们通过在一个物种内的多个个体上测试引物，然后应用已建立的计算工具，展示了一种为给定物种选择最佳引物的方法。这些参考引物可以促进任何具有生态或经济利益的脊椎动物物种中基于 qPCR 的端粒长度测量。