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Sampling times are often used for calibration since some organisms accumulate enough mutations over the course of their sampling period. This practice ties together two key concepts: measurably evolving populations and the phylodynamic threshold. Our current understanding suggests that populations meeting these criteria are suitable for molecular clock calibration via sampling times. However, the definitions and implications of these concepts remain unclear. Using Hepatitis B virus-like simulations and analyses of empirical data, this study shows that determining whether a population is measurably evolving or has reached the phylodynamic threshold does not only depend on the data, but also on model assumptions and sampling strategies. In Bayesian applications, a lack of temporal signal due to a narrow sampling window results in a prior that is overly informative relative to the data, such that a prior that is potentially misleading typically requires a wider sampling window than one that is reasonable. In our analyses we demonstrate that assessing prior sensitivity is more important than the outcome of tests of temporal signal. Our results offer guidelines to improve molecular clock inferences and highlight limitations in molecular sequence sampling procedures.<\/jats:p>","DOI":"10.1093\/molbev\/msag133","type":"journal-article","created":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T11:39:32Z","timestamp":1780141172000},"source":"Crossref","is-referenced-by-count":0,"title":["The Phylodynamic Threshold of Measurably Evolving Populations"],"prefix":"10.1093","volume":"43","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-7854-2121","authenticated-orcid":false,"given":"Ariane","family":"Weber","sequence":"first","affiliation":[{"name":"Human Palaeosystems Research Group, Max Planck Institute of Geoanthropology , Jena ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7287-9582","authenticated-orcid":false,"given":"Julia","family":"Kende","sequence":"additional","affiliation":[{"name":"Bioinformatics and Biostatistics Hub, Institut 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