This study examined differences in classroom climate perceptions and motivational beliefs between students enrolled in undergraduate Biology courses that fostered an active learning environment through pedagogical enhancements (intervention group) and students enrolled in traditional, exposition-centered Biology courses (control group). Participants were 962 students who self-selected into one of the introductory biology courses randomly assigned to the intervention or control. Multiple regression analyses indicated that students in classrooms receiving the intervention held higher self-efficacy, personal value, and interest for the course at completion compared to their counterparts in traditional classrooms. Moreover, students receiving the intervention reported higher levels of instructor support, a perception of higher expectations for learning and understanding (academic press), and a greater feeling that the course was interesting and valuable (situational interest). Among students enrolled in the intervention, perceptions about the value of pedagogical enhancements (e.g., clicker technology) were associated with positive perceptions of the classroom climate and adaptive motivational beliefs. Results gained from this intervention may be broadly useful to other campuses interested in increasing student success even when faced with large enrollments and minimal faculty and staff support.