UW-Stout/Growth Curve

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  1. Three days before the experiment, streak knockout strains onto YPD plates.
  2. The morning before the experiment, pick a colony from each strain into 5 ml YPD broth in a disposable test-tube. Incubate on the roller drum at 30°C for 8-10 hours.
  3. The evening before the experiment, transfer 20 ul of each strain into 5 ml of fresh YPD broth. Incubate on a roller drum at 30°C overnight.
  4. The morning of the experiment, dilute each culture to an OD600 of 0.1-0.2 in YPD broth. Return to the drum roller until ready to test.
  5. Transfer 50 µl of yeast culture and 50 µl of sterile water to a well in the assay plate.
  6. Set up the plate reader as follows:
    • Temperature: 30°C
    • Mode: Kinetic
    • Read: 600 nm
    • Interval: 5 minutes
    • Total run time: 24 hours
    • Shake before read: 30 seconds
  7. Transfer the assay plate to the reader and run for 24 hours.