Interhemispheric laterality has often been linked to different behavioural styles. This study investigates the link between limb preference and personality in donkeys. The sample consisted of 47 donkeys (Equus asinus), 30 males and 17 females. Limb preference was determined using observation of the leading limb in a motionless posture and personality was measured using the Donkey Temperament Questionnaire (French, J. M. (1993). Assessment of donkey temperament and the influence of home environment. Applied Animal Behaviour Science, 36(2), 249–257. doi:10.1016/0168-1591(93)90014-G) completed by the donkeys’ keepers. A Principal Component Analysis obtained two components: Agreeableness and Extraversion. Age showed a positive relationship with Agreeableness, echoing trends in humans Donkeys did not show a population-level preference towards either side. Limb preference significantly predicted the trait difficult to handle: donkeys with a preference to keep the right foot forward when motionless were harder to handle. This study presents the first investigation into limb preference and personality in donkeys, although more research is needed to clarify whether there is a population-level limb preference bias in donkeys, and the relationship between limb preference and Agreeableness.