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: