Artificial control of mating type and repeated mating to produce polyploid cells in Saccharomyces cerevisiae

The budding yeast Saccharomyces cerevisiae is an excellent model for examining the effects of ploidy. Here, we provide a protocol for producing polyploid cells by creating a basic unit (matΔ) and polyploidizing it via repeated mating. We describe steps for basic unit construction by one-step transformation, increased ploidy via repeated mating, and ploidy confirmation using flow cytometry. This protocol can be broadly applied to evaluate the physiology of polyploid cells. For complete details on the use and execution of this protocol, please refer to Oya and Matsuura (2022).¹.