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In order to discern sake yeast, shochu yeast, and wine yeast, a specific polymerase chain reaction (PCR) was investigated. PCR products between shochu yeast strains and sake yeast weredistinguished by using a primer designed with an open reading frame of FLO5. Wine yeasts and most of other yeasts were distinguished by a primer designed with an open reading frame of YHR213W.However, PCR products of shochu yeast KF 1 resembled those of some wine yeasts, and we couldnot discern them. As this method based on the PCR to discern brewing yeasts was simpler than classical physiological methods, it was useful to discern brewing yeast with different purposes for application.