Difference between revisions of "UW-Stout/Heat Shock SP21"
| Line 4: | Line 4: | ||
'''Calibration Experiment Materials''': | '''Calibration Experiment Materials''': | ||
• Distilled H2O | • Distilled H2O | ||
| + | |||
• Wild-type yeast cells | • Wild-type yeast cells | ||
| + | |||
• P20 and P200 micro pipettors | • P20 and P200 micro pipettors | ||
| + | |||
• PCR test tubes (mini) | • PCR test tubes (mini) | ||
| + | |||
• Glass beads | • Glass beads | ||
| + | |||
• 15 agar filled petri dishes | • 15 agar filled petri dishes | ||
Revision as of 10:31, 27 April 2021
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:
