Objective This study aimed to investigate allied health professionals' (AHPs') perspectives pre- and post-implementation of an electronic medical record (EMR) in a tertiary health service in Australia and examine factors influencing user acceptance. Methods Data were collected pre- and post-EMR implementation via cross-sectional online surveys based on the Unified Theory of Acceptance and Usage of Technology (UTAUT). All AHPs at a large tertiary hospital were invited to complete the surveys. Data analysis included descriptive analysis, Mann-Whitney U tests for pre-post item- and construct-level comparison and content analysis of free-text responses. The theoretical model was empirically tested using partial least squares structural equation modelling. Results AHPs had positive attitudes toward EMR use both pre- and post-implementation. Compared to pre-implementation, AHPs felt more positive post-implementation about system ease of use and demonstrated decreased anxiety and apprehension regarding EMR use. AHPs felt they had adequate resources and knowledge to use EMR and reported real-time data accessibility as a main advantage. Disadvantages of EMR included an unfriendly user interface, system outages and decreased efficiency. Conclusions As AHPs increase EMR system familiarity, their positivity towards its use increases. An understanding of what influences AHPs when implementing new compulsory technology can inform change management strategies to improve adoption.