Difference between revisions of "UW-Stout/Heat Shock SP21"
(Created page with "'''Preliminaries''': None '''Calibration Experiment Materials''': • Distilled H2O • Wild-type yeast cells • P20 and P200 micro pipettors • PCR test tubes (mini) •...") |
|||
| Line 13: | Line 13: | ||
1. Vortex the yeast culture briefly to resuspend the yeast pellet. | 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. | 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”. | 3. Label each tube “A, B, C, D, E”. | ||
| + | |||
4. Insert test tube A in the thermocycler for exactly 30 minutes at 48°C. | 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. | 5. After test tube A is done cycling, place test tube B in the thermal cycler for exactly 15 minutes at 48° C. | ||
| Line 22: | Line 25: | ||
9. Carry on and analyze/count the DNA colonies in the wells. | 9. Carry on and analyze/count the DNA colonies in the wells. | ||
| − | + | ||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
| − | |||
'''Knockout Experiment Materials''': | '''Knockout Experiment Materials''': | ||
Revision as of 10:30, 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:
