In response to calls for reform in undergraduate biology education, we conducted research examining how varying active-learning strategies impacted students’ conceptual understanding, attitudes, and motivation in two sections of a large-lecture introductory cell and molecular biology course. Using a quasi-experimental design, we collected quantitative data to compare participants’ conceptual understanding, attitudes, and motivation in the biological sciences across two contexts that employed different active-learning strategies and that were facilitated by unique instructors. Students participated in either graphic organizer/worksheet activities or clicker-based case studies. After controlling for demographic and presemester affective differences, we found that students in both active-learning environments displayed similar and significant learning gains. In terms of attitudinal and motivational data, significant differences were observed for two attitudinal measures. Specifically, those students who had participated in graphic organizer/worksheet activities demonstrated more expert-like attitudes related to their enjoyment of biology and ability to make real-world connections. However, all motivational and most attitudinal data were not significantly different between the students in the two learning environments. These data reinforce the notion that active learning is associated with conceptual change and suggests that more research is needed to examine the differential effects of varying active- learning strategies on students’ attitudes and motivation in the domain.