In instructional settings, evolution and natural selection are challenging concepts to teach, due to the fact that these topics are difficult to observe in the laboratory or lecture hall. In the past few years, Avida-ED has emerged as an innovative tool for teaching evolutionary principles. It allows students to directly observe effects of evolution by changing different variables, such as environmental conditions and genetic sequences. In our study, we used pretest and posttest questionnaires to investigate the use of Avida-ED in undergraduate coursework. We showed that students demonstrated similar improvement in evolutionary understanding irrespective of major, undergraduate level, sex, or final end-of-course grade. These results reinforce the idea that Avida-ED can facilitate learning of evolution in all student populations.