Purpose: Nurses lead Evidence-Based Practice (EBP) in the healthcare system. To facilitate the evidence-based nursing process, nurses must have the knowledge and skills to formulate clinical questions in a searchable format. The purpose of this study was to develop an e-learning program on steps of EBP, especially focusing on clinical questioning to assess its effects. Methods: This research utilized a quasi-experimental study with a nonequivalent control group pre-post test design. The experimental group was provided an e-learning program with case-based animation. The control group was provided written material about EBP. The e-learning program was performed from October to November 2015. To evaluate the effects of the program, knowledge of EBP, attitude towards EBP, practice of EBP, clinical questioning confidence, and clinical questioning knowledge were evaluated. Results: There were significant differences in attitude towards EBP (t=2.08, p=.042), practice of EBP (t=2.06, p=.044), clinical questioning confidence (t=2.05, p=.045) and clinical questioning knowledge (t=2.08, p=.042). However, there was no significant difference between the experimental and control groups in knowledge of EBP (t=1.20, p=.237). Conclusion: The e-learning program with case-based animation was an effective method to provide the EBP education to nurses. Further researches are needed to disseminate the case-based education to nurses and nursing students and to develop more diverse educational media. [ABSTRACT FROM AUTHOR]