The current boom of dam construction at low latitudes endangers the integrity and function of major tropical river systems. A deeper understanding of the physical and chemical functioning of tropical reservoirs is essential to mitigate dam-related impacts. However, the development of predictive tools is hampered by a lack of consistent data on physical mixing and biogeochemistry of tropical reservoirs. In this study, we focus on Lake Kariba (Southern Africa), the largest artificial lake in the world by volume. Kariba Dam forms a transboundary reservoir between Zambia and Zimbabwe, and therefore its management represents a socio-politically sensitive issue because the Kariba Dam operation completely changed the downstream hydrological regime. Although Lake Kariba represents a unique and scientifically interesting case study, there is no consistent dataset documenting its physical and chemical behaviour over time. This limits the scope for quantitative studies of this reservoir and its downstream impacts. To address this research gap, we aggregated a consistent database of in situ measurements of temperature and oxygen depth profiles for the entire 60 years of Lake Kariba’s lifetime and performed a detailed statistical analysis of the thermal and oxygen regime of the artificial lake to classify the different behaviours of the lake’s sub-basins. We demonstrate that the seasonal stratification strongly depends on the depth of the water column and on the distance from the lake inflow. Satellite data confirm these spatiotemporal variations in surface temperature, and reveal a consistent longitudinal warming trend of the lake surface water temperature of about 1.5°C from the inflow to the dam. Finally, our results suggest that the stratification dynamics of the lacustrine sub-basins have the potential to alter the downstream Zambezi water quality. Future research should focus on assessing such alterations and developing strategies to mitigate them.