All Canadian lake sturgeon (Acipenser fulvescens) populations are listed under the Committee on the Status of Endangered Wildlife in Canada (COSEWIC) due to their complex life history, loss of habitat and negative interactions with anthropomorphic activities. Lake sturgeon diets vary considerably across their range and depend on the local benthic invertebrate fauna, substrata, and competition with congener species. Hence, determining population-specific prey assemblages is a vital contribution to effective conservation efforts. We used carbon and nitrogen stable isotopes to identify lake sturgeon prey preferences for juvenile (<100 cm) and adult (>100 cm) age classes and their trophic niche in the Saskatchewan River, SK, Canada. In this system, lake sturgeon were the top predator within the benthic food web with no direct competition for preferred prey items from congener species. Their diet was dominated by crayfish (49.1± 6.4%) and snails (36.3 ± 5.5%), with no significant differences between age classes. Furthermore, lake sturgeon favoured deep rocky pools throughout the river; a habitat where crayfish and snails are typically found. Therefore, conservation efforts should be directed at preserving these habitats for the residing benthic invertebrate community, and the lake sturgeon's ability to access them.