UW-Stout/Ultraviolet Light

Materials

• YPD media, both broth and 2% agar plates]
• Glycerol stocks of knockout strains
• Cell 60 mm Culture Dish, containing 6 ml of agar

Equipment

• Incubator set to 30°C.
• Bachur & Associates Sanata Clara, CA 95050 Model LS-100-3 UV Light Exposeure System

Protocol

1. Three days before the experiment, streak knockout strains onto YPD plates.
2. One day 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 overnight.
3. The morning of the experiment, dilute each culture to an OD600 of 0.2 in YPD broth. Return to the drum roller for between 30 and 90 minutes.
4. Transfer 33 µl to a well in the Culture Dish.
5. Set up the UV light Exposure System as follows:
   • 400 watts
   • time increments
   • place in the incubator in dark for 48 hours