UW-Stout/UV Light SP21

Materials

• Cell 60 mm Culture Dish, containing 6 ml of agar
• Phosphate Buffered Saline (PBS)
• Glycerol stocks of yeast strains

Equipment

• Incubator set to 30°C.
• Bachur & Associates Sanata Clara, CA 95050 Model LS-100-3 UV Light Exposeure System
• P1000 and P10 Micro-pipettes

Calibration Protocol

2. Vortex yeast stock to resuspend the yeast cells.
3. Pipette 10ul of yeast stock into the 9,990ul of PBS to create a dilution containing 2 yeast cells per microliter.
4. Prepare 6 plates with 50ul of the wild yeast dilution.
5. Label each plate individually 500, 600, 700, 800, 900 and 1000 for the number of seconds each plate will be exposed to the UV light.
6. Set up the UV light exposure system:

**400 watts

• • desired time increment

1. Run yeast plates without top under the UV light for their respective times in seconds.
2. Place plates upside down in dark incubator set to 30°C for 48 hours.