AbstractNatural radiogenic isotopes (primarily 87Sr/86Sr) from hot springs in the Upper Geyser Basin of the Yellowstone Plateau volcanic field and associated rocks were used to evaluate groundwater flow patterns, water‐rock reactions, and the extent of mixing between various groundwater sources. Thermal waters have very low uranium concentrations and 234U/238U activity ratios near 1.0, which limit their utility as tracers in this reducing setting. Thermal waters have higher Sr concentrations (<22 ng/g) and a wide range of 87Sr/86Sr values that vary both temporally at individual discharge sites and between adjacent springs, indicating that conduits tap different subsurface reservoirs to varying degrees. Sr from local rhyolites have 87Sr/86Sr compositions that bound the range of values observed in groundwater throughout the basin. Non‐boiling springs on the west flank of the basin discharge water with low 87Sr/86Sr consistent with flow through young volcanic rocks exposed at the surface. Boiling springs in the central basin have higher 87Sr/86Sr values reflecting interactions with older, more radiogenic volcanic rocks. Variability in upwelling thermal waters requires mixing with a low 87Sr/86Sr component derived from young lava or glacial sediments, or more likely, from deeper sources of hot groundwater circulating through buried Lava Creek Tuff having intermediate 87Sr/86Sr. Isotope data constrain basin‐wide output of thermal water to 110–140 kg·s−1. Results underscore the utility of radiogenic Sr isotopes as valuable tracers of hydrothermal flow patterns and improve the understanding of temperature‐dependent water‐rock reactions in one of the largest continental hydrothermal systems on Earth.