Objective: The present study examined potential barriers to cycle-based transport amongst undergraduate students, to inform the design of future cycling promotion interventions at the University of Johannesburg (UJ). Participants: A total of 606 first, second and third year UJ undergraduates took part. Methods: Sociodemographic and economic determinants of bicycle/car ownership, cycling competency and behavior were evaluated using data derived from a 9-item questionnaire, before and after adjustment for measured confounders. Results: While 70% of respondents knew how to cycle, only 26% owned/had access to a bicycle, and only 18% had last cycled for transport. Bicycle ownership and competency were far commoner amongst male and older participants, and those whose parents had the means to own a car. Conclusions: Interventions to promote cycle-based transport must address the many (predominantly female) students who: have limited cycling competency; do not own/have access to a bicycle; or have little/no experience of cycling for transport.