Background: In the literature, there is increasing interest in the effects of subliminal stimuli on humans. Objectives: To investigate the effects of the positive subliminal messages placed in a video used for ventrogluteal intramuscular injection training of nursing students on students’ remembering, anxiety, self-confidence, general self-efficacy and practice skill levels. Methods: The population of the randomized controlled study conducted between 9-10 October 2018 consisted of first year nursing students at a faculty of health sciences (n=64). An animated video on ventrogluteal intramuscular injection that contained subliminal messages was shown to the students in the experiment group. In the study, the students in the control group were given only practice together with the theoretical education on the subject. A Student Information Form, Trait-State Anxiety Scale, General Self-Efficacy Scale, Self-Confidence Scale, and Ventrogluteal Intramuscular Injection Practice Skill Evaluation Form were used. In the study, the independence Chi-square test was used to compare the experiment and control groups. Results: Although statistically non-significant (p>0.05), identifying ventrogluteal intramuscular injection sites (n=21, 33.9%) and proper injection technique (n=22, 35.5%) steps were remembered more by students in the experiment group. It was found that ventrogluteal intramuscular injection practice skill was at ‘adequate’ level in the experiment group (n=21, 35%). Conclusion: Visual subliminal messages used in ventrogluteal intramuscular injection training have no effect on nursing students’ anxiety, self-confidence, and general self-efficacy levels. Basic nursing skill applications can be prepared with animated videos in a way to attract students’ attention.