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Abstract The resources for carrying out and analyzing microbial evolution experiments have become more accessible, making it possible to expand these studies beyond the research laboratory and into the classroom. We developed five connected, standards‐aligned yeast evolution laboratory modules, called “yEvo,” for high school students. The modules enable students to take agency in answering open‐ended research questions. In Module 1, students evolve baker's yeast to tolerate an antifungal drug, and in subsequent modules, investigate how evolved yeasts adapted to this stressful condition at both the phenotype and genotype levels. We used pre‐ and post‐surveys from 72 students at two different schools and post‐interviews with students and teachers to assess our program goals and guide module improvement over 3 years. We measured changes in student conceptions, confidence in scientific practices, and interest in STEM careers. Students who participated in yEvo showed improvements in understanding of activity‐specific concepts and reported increased confidence in designing a valid biology experiment. Student experimental data replicated literature findings and has led to new insights into antifungal resistance. The modules and provided materials, alongside “proof of concept” evaluation metrics, will serve as a model for other university researchers and K − 16 classrooms interested in engaging in open‐ended research questions using yeast as a model system.