-URA Selection

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