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Spores are essential for the long-term survival of many diverse organisms, due to their roles in reproduction and stress resistance. In the environmental human fungal pathogen, <i>Cryptococcus</i>, basidiospores are robust cells with the ability to cause disease in animal models of infection. Here we describe methods for producing and purifying <i>Cryptococcus</i> basidiospores in quantities sufficient for large-scale analyses. The production of high numbers of pure spores has facilitated the development of new assays, including quantitative germination assays, and enabled transcriptomic, proteomic, and virulence studies, leading to discoveries of behaviors and properties unique to spores and spore-mediated disease.