Difference between revisions of "UW-Stout/UV Light SP21"
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#Vortex yeast stock to resuspend the yeast cells. | #Vortex yeast stock to resuspend the yeast cells. | ||
#Pipette 10ul of yeast stock into the 9,990ul of PBS to create a dilution containing 2 yeast cells per microliter. | #Pipette 10ul of yeast stock into the 9,990ul of PBS to create a dilution containing 2 yeast cells per microliter. | ||
− | # | + | #Vortex dilution and prepare 6 plates with 50ul of the wild yeast dilution. |
#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. | #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. | ||
#Set up the UV light exposure system: | #Set up the UV light exposure system: |
Revision as of 15:43, 26 April 2021
Contents
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
- Vortex yeast stock to resuspend the yeast cells.
- Pipette 10ul of yeast stock into the 9,990ul of PBS to create a dilution containing 2 yeast cells per microliter.
- Vortex dilution and prepare 6 plates with 50ul of the wild yeast dilution.
- 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.
- Set up the UV light exposure system:
- 400 watts
- desired time increment
- Run yeast plates without top under the UV light for their respective times in seconds.
- Place plates upside down in dark incubator set to 30°C for 48 hours.