BACKGROUND: Undergraduate involvement in research necessitates better understanding of their potential, as well as the anticipated barriers they will face. The objective of this study was to assess medical undergraduates' knowledge, attitudes, and research barriers. METHODS: This was a cross-sectional study conducted on medical students from the University of Tripoli, Libya, where a self-administered questionnaire was used to evaluate the students' knowledge, attitudes, and perceived barriers. Filled questionnaires were received from the medical undergraduates of different faculties of medical specialties. Data were analyzed using descriptive statistics. RESULTS: A total of completed questionnaires were received from 120 undergraduate medical students. The overall result of knowledge among them was encouragingly good (51%). Majority of the students taught that the main barriers in conducting research were lack of awareness (71.7%), lack of self-interest (56.7%), lack of faculty encouragement for research (84.2%), insufficient time (59.2%), and difficulty in obtaining resources and data for research (68.3%). CONCLUSION: Participants in the current study showed a moderate knowledge level with associated positive attitudes toward research. This attitude needs to be transformed into better knowledge and appropriate practice.