Abstract Background The evolution education research community has defined the construct of “evolution acceptance” in different ways and measured it using different instruments. One of these instruments—the GAENE—has not been analyzed across different student populations, demographic groups, degree plans, and instructional treatments. Such comparisons are crucial for examining whether the inferences drawn from instrument measures are valid, reliable, and generalizable. In this study, we attempt to replicate findings produced in the original validation study and explore aspects of the instrument not previously examined. Methods We use Rasch analysis to study a large sample (n > 700) of undergraduates enrolled in standard introductory biology...