This mixed methods study explores college biology students' perceptions of the utility of evolution knowledge. In Phase I, 14 college biology students from an upper-level Evolution course were interviewed about their evolution acceptance and perceptions of the utility of evolutionary knowledge. In Phase II, findings from Phase I were used to develop a 28-item quantitative Usefulness of Evolution Knowledge Survey. Then, 472 college biology students enrolled in three biology courses were given the Usefulness of Evolution Knowledge survey to identify (1) evolution topics that have the highest utility value for evolution learners, (2) careers for which evolution knowledge is deemed most useful, and (3) relationships among evolution utility value and other constructs. The most useful evolutionary topics were genetic variation and adaptations of particular organisms to particular environments. Evolutionary knowledge was deemed most useful for conservation careers, with less utility for medical and forensic careers, and far less utility for careers in physics or chemistry. Significant relationships were found between perceptions of the usefulness of evolution and evolution acceptance, understanding of evolution, and intended career. Findings are discussed within the framework of the Expectancy Value Theory, and implications for future pedagogical interventions and associated research are provided.