YPD medium (2% YPED)

1. To an autoclaveable flask add:

- 10g/L Yeast Extract

- 20g/L Bacto Peptone

- ddH₂O to 1L

- For plates: 20g/L agar

2. Mix well and autoclave

3. Add:

- 20g/L Glucose (to conserve filters, preferable 50mL of 40% w/v glucose filtered after autoclave)

Synthetic Drop-out medium

4. Το an autoclaveable flask add:

- 1.39-1.92g/L Yeast Synthetic Drop-out medium (depends on the medium)

- 6.7g/L Yeast Nitrogen Base without amino acids

- For plates: 20g/L agar

5. Mix well and autoclave

6. Add:

- 20g/L Glucose (to conserve filters, preferable 50mL of 40% w/v glucose filtered after autoclave)

Pre-sporulation medium (1% YPA)

7. To an autoclaveable flask add:

- 10g/L Yeast Extract

- 20g/L Peptone

- 10g/L Potassium Acetate

- For plates: 20g/L agar

8. Mix well and autoclave

Sporulation medium

9. To an autoclaveable flask add:

- 10g/L Potassium Acetate

- For plates: 20g/L agar

10. Mix well and autoclave

6% YEPD

11. To an autoclaveable flask add:

- 10g/L Yeast Extract

- 20g/L Bacto Peptone

- ddH₂O to 1L (850mL)

- For plates: 20g/L agar

12. Mix well and autoclave

13. Add:

- 60g/L Glucose (To conserve filters, preferable 150mL of 40% w/v glucose filtered after autoclaving)

Nitrogen-starvation medium (SD(–N) medium)

14. Το an autoclaveable flask add:

- 6.7g/L Yeast nitrogen base without amino acids and ammonium sulfate

15. Mix well and autoclave

16. Add:

- 20g/L Glucose (to conserve filters, preferable 50mL of 40% w/v glucose filtered after autoclave)

Preparing plates

17. Aliquot flask into a single 50mL falcon tube when the flask is at a temperature which feels comfortable to hold for a prolonged period of time (~55 ^oC)

18. Use every 50mL Falcon for approximately 3 plates.

Preparing filtered glucose (optional)

19. If glucose fails to dissolve in the initial flask, aliquot in a 50mL falcon and use the vortex machine and water bath to slowly heat up the solution and allow the glucose to dissolve.

20. Also make sure to add the glucose in small quantities in the initial flask.