This thesis deals with possibilities of isolation DNA yeasts genus Brettanomyces from wine with following identification using PCR. Fourteen wine samples with assumption of occurrence Brettanomyces were analysed. The literary section describes characteristic of yeasts Brettanomyces and their impact on wine. Further is described principle of PCR and use of this method in detection of Brettanomyces. Four different methods of isolation DNA directly from wine were tested. Wine samples were analysed using of five different primers by PCR. In this work were tested designed probe and primers for identification of Brettanomyces by real-time PCR.