Using current best evidence to make decisions can improve outcomes of public health. Thus, establishing the capacities of evidence-based public health (EBPH) has become one of the core competences. To better scale up EBPH movement, efforts should focus on introduction of EBPH into school curriculum. However, data indicating the extent to which EBPH is used in universities are scant. In the current study, we conducted a nationwide questionnaire survey to investigate the perceptions toward EBPH among full-time teachers and senior undergraduates at all universities with a Department of Public Health in Taiwan. A structured questionnaire was distributed by post to all potential participants of nine universities in 2017. Questions included items related to awareness, knowledge, skills, behaviors, and barriers of EBPH. Results showed teachers were more aware of EBPH than students. In addition, teachers more often had sufficient knowledge and skills of EBPH, and more often applied the findings to decision-making after critical appraisal than students. Furthermore, personal barriers toward EBPH were more common in students than teachers. In conclusion, there are differences in awareness, knowledge, skills, behaviors, and barriers of EBPH between teachers and students. The data suggest that an initiative of systematically teaching EBPH to undergraduates is important.