The purpose of this study was to determine the effect of teaching with animation on chemistry achievement and learning persistence on pre-service science teachers studying in the undergraduate program of science teaching. The quasi-experimental model, one of the quantitative research approaches, was used in the study. The pretest-posttest was applied with the experimental control group. Eighty-two pre-service teachers, randomly selected with the appropriate sampling method, participated in the research. The "General Chemistry Achievement Test" developed by Sönmez (2017), which has validity and reliability in the literature, was used as a data collection tool. In addition, to determine the persistence of learning, chemistry achievement test post-test applications were applied again at six-week intervals. The data obtained from the findings was analyzed using descriptive and inferential statistical techniques in the SPSS 23.0 package program. Because the collected data did not have a normal distribution, the research conducted using nonparametric test techniques (Mann Whitney U and Kruskal Wallis) revealed a significant positive difference in favor of the experimental group to which animation was applied. In addition, in terms of gender variables, it has been determined that women have a higher mean than men. In contrast, there was no significant difference in grade level between teacher candidates. Following a six-week persistence test, it was found that there was a significant and positive correlation, ensuring the learning's persistence.