Background: There are deficiencies in ensuring the permanence of some theoretical information taught in nursing education and transferring it to practice environment. Mobile-assisted teaching can be useful to eliminate deficiencies. The aim of this study was to determine the effect of mobile-assisted teaching on nursing students' learning ventrogluteal injection. Methodology: The study was conducted in Turkey between February and June 2022. This study is a single group pre-posttest intervention. The study sample consisted of 354 students studying in the 2nd, 3rd and 4th grades in Turkey. After the students completed the Introductory Characteristics Form and Ventrogluteal Region Information Suggestion Form in the pre-test, the researchers sent a ventrogluteal injection animation video to their mobile phones. The students who watched the video completed the Ventrogluteal Region Information Suggestion Form and Mobile Education Activity Form in the final test. Results and Conclusion: There was a statistically significant difference between the pre-posttest score medians of the students (p < 0.001). While the preference of the students for the ventrogluteal region in intramuscular injection was 28.5% before mobile learning, it increased to 51.1% after the training. In this study, after the training given through mobile learning, the knowledge level of the students about ventrogluteal injection and their preferences for ventrogluteal injection increased. In line with these results, mobile-assisted education should be used in nursing education.