Sporulation of Budding Yeast:

Starvation of diploid yeast cells for nitrogen and carbon sources induces meiosis and spore formation. The sporulation process can be induced in cells growing either on solid or in liquid medium.

1. Sporulation in liquid media:
2. Grow the diploid to be sporulated to an OD600 of 2.5 to 3.0 in YPD medium.
3. Transfer 1 ml culture to a sterile, disposable 15-ml tube and centrifuge 5min at 1200xg (3000rpm).
4. Pour off supernatant and resuspend cells in 5ml sterile water. Vortex to resuspend cells and repeat spin.
5. Pour off supernatant and resuspend cells in 1ml liquid sporulation medium supplemented with nutritional requirements of the particular diploid.
6. Shake at 30⁰C for 3-6 days.
7. Look for sporulation microscopically.

Sporulation Medium:

10 g/l potassium acetate

1 g/l yeast extract

0.5 g/l dextrose