Yeast Mating with Diploids Isolation

Yeast Mating with diploid Isolation

Materials

• YPD plates

Method

• In the evening inoculate a small amount of the two strains to be mated in 5ml YPD;
• Incubate static at 30°C overnight;
• Resuspend completely the cells and dilute 1:2000 in sterile water;
• Plate 100 microl on a YPD plate and incubate overnight at 30°C;
• Look for tiny colonies: pick and patch ~10 putative diploids — diploids colonies are generally bigger: start with the biggest colonies;
• Check ploidy by FACS and/or sporification assay;