Springer Lab: Sporulation
1.Take an overnight YEPD saturated cultures(~10^8 cells/mL) Dilute 1:100, let grow for 4 hours at 30 C.
2.Take 1ml and wash once in water and then resuspend in 2 mL SPO media.
Sporulation media SPO
- Potassium acetate - 10g
- Bacto-yeast extract - 1g
- Glucose - 0.5g
- H20 - 1L
3.Let the cultures sit overnight at room temperature and then incubate at 30C with shaking for several days.
Note: More dense cultures lead to accumulation of bicarbonate and a higher sporulation efficiency.
Also for low efficiency sporulants (petite, or mitochondria mutations) carbonate can be added to change the pH and increase the sporulation efficiency. For more details check Ohkuni and Hayashi. Bicarbonate‐mediated social communication stimulates meiosis and sporulation of Saccharomyces cerevisiae. Yeast (1998).