Background: Fostering pre-service teachers' acceptance of evolutionary theory and their preference for its teaching implies knowledge of the factors which influence both constructs. This study aims to explore how cognitive (knowledge of evolution), affective (attitude towards religion and science, scientism, and creationism), and contextual factors (age, gender, parents' educational qualification, semester, teacher education program) are related to acceptance and preference. Furthermore, the study aims at exploring the relationship between acceptance and preference. Methods: A total of 180 German pre-service biology teachers participated in the study.