The Barstow Formation in the Mojave region of California was deposited in an extensional-basin setting of the Basin and Range province and preserves diverse middle Miocene mammalian assemblages. Six facies associations represent the dominant depositional environments in the basin, which changed through time from alluvial-fan and playa-dominated settings to floodplains and spring-fed wetlands. The majority of fossil localities and specimens occur in later-forming facies associations. We analyzed the taphonomic characteristics of fossil assemblages to test whether basin-scale facies associations or locality-scale facies exert more control on the preservational features of mammalian assemblages through the formation. We documented the facies settings of 47 vertebrate localities in the field in order to interpret depositional setting and the mode of accumulation for fossil assemblages. We evaluated skeletal material in museum collections for taphonomic indicators, including weathering stage, original bone-damage patterns, hydraulic equivalence, and skeletal-element composition. We evaluated four alternative modes of accumulation, including attritional accumulation on the land surface, accumulation by fluvial processes, carnivore or scavenger accumulations, and mass-death events. The majority of localities represent attritional accumulations at sites of long-term mortality in channel-margin, abandoned-channel, poorly drained floodplain, and ephemeral-wetland settings. Skeletal-element composition and taphonomic characteristics varied among facies, indicating an important role for depositional setting and landscape position on fossil-assemblage preservation. We find that locality-scale facies have a greater influence on the taphonomic characteristics of fossil assemblages; the taphonomy of each facies association is influenced by the facies that compose it. The facies composition and distribution within facies associations change through the formation, with a greater variety of depositional settings forming later in the history of the basin. Heterogeneous landscapes present more settings for fossil accumulation, contributing to the increase in fossil occurrence through the depositional history of the formation.