Protocols &Technical Information
Yeast Deletion Database (consortium members only)
Sporulation Method for BY4743.v1
click here for v.2
Efficient sporulation of BY4743 requires growth as a patch on freshly prepared GNA presporulation plates at 30°C overnight prior to transfer to liquid sporulation medium. (Note: if GNA is inoculated with an old culture, transfer to another GNA plate for an additional day.) Resuspend one colony equivalent of cells into 2mls of liquid sporulation medium in a 17x150mm glass test tube. Sporulation cultures should be incubated on a roller wheel for 3-5 days at 25°C. The cultures should be ready to dissect and can be stored at 4°C for a couple of weeks. However, dissection within two weeks is the best. Sporulation efficiencies should be around 20-30%. If you prespore on YPD instead of GNA the sporulation efficiency is much lower.
GNA Presporulation Plates
Final Concentrations recipe for 2 plates recipe for 10 plates 5% D-glucose 3% Difco nutrient broth 1% Difco yeast extract 2% Bacto agar Autoclave for 15 minutes. Plates should always be freshly made and last only for 1-2 weeks at 4°C for optimum sporulation.
Liquid Sporulation Medium
Final Concentrations Stock solutions 1% Potassium Acetate 10% Potassium Acetate (20g/200ml) 0.005% Zinc Acetate 0.5% Zinc acetate (1g/200ml) 2 mls
Resuspend the stocks in dH2O and filter sterilize. The stocks are mixed in sterile water just before sporulation.