UW-Stout/Heat Shock SP21
Preliminaries: None
Calibration Experiment Materials: • Distilled H2O • Wild-type yeast cells • P20 and P200 micro pipettors • PCR test tubes (mini) • Glass beads • 15 agar filled petri dishes
Calibration Experiment Procedure:
1. Vortex the yeast culture briefly to resuspend the yeast pellet.
2. Set up 5 different PCR test tubes with 50 ul of wild-strain yeast cells and 50 ul of distilled water in each.
3. Label each tube “A, B, C, D, E”.
4. Insert test tube A in the thermocycler for exactly 30 minutes at 48°C. 5. After test tube A is done cycling, place test tube B in the thermal cycler for exactly 15 minutes at 48° C. 6. After test tube B is done cycling, place test tube C in the thermal cycler for exactly 30 minutes at 48° C. 7. After test tube C is done cycling, place test tube D in the thermal cycler for exactly 45 minutes at 48° C. 8. After test tube D is done cycling, place test tube E in the thermal cycler for exactly 60 minutes at 48° C. 9. Carry on and analyze/count the DNA colonies in the wells.
Knockout Experiment Materials: