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- K. marxianus is grown overnight in 3ml of YPD media.
- Centrifuge the culture tubes at max rpm and wash once with sterile distilled water and centrifuge again to get rid of the wash.
- Resuspended in 1ml of sterile distilled water.
- Aliquot 150ul of cells.
- Spread the cell aliquots on 5-FOA plates and expose the plates to UV light for 20 seconds.
- Incubate plates at 30°C for 2 days. Pick the colonies.
- Picked colonies are grown in minimal media with and without uracil and observerd for growth with uracil.