UW-Stout/Copper FA21

Materials & Equipment

• CuSO4
• Sterile Water
• 1000ul Pipette
• 50ul Pipette
• Centrifuge Tubes
• Wild Type Yeast Cells
• Yeast Cells with Your Favorite Gene removed.
• Vortex
• Disposable Pipette Tips
• Plate Reader
• Wells

Personal Protective Equipment

• Latex Gloves
• Eye Pro
• Lab Coat

Wild Type Protocol

1. Vortex Wild type Yeast Solutions
2. Create Solutions with Copper Sulfate and Sterile water at the following Concentrations.
   1. 20mM (Solution) Copper Sulfate
   2. 10mM Copper Sulfate
   3. 5.0mM Copper Sulfate
   4. 2.0mM Copper Sulfate
   5. 1.0mM Copper Sulfate
   6. 0.5mM Copper Sulfate
   7. 0.2mM Copper Sulfate
   8. 0.1mM Copper Sulfate
3. Combine 50ul of the Copper Sulfate solution and 50ul of the wild type yeast cells and put them in eight different wells.
4. Take the wells that are prepared with the yeast and the Copper Sulfate and put it in a plate reader and leave it there for 24 hours.

Protocol with Gene Removed

Part 1

1. Insert 50ul of yeast cells in different wells.
2. Insert 50ul of CuSO4 at 0.5mM concentration into the same wells as the yeast cells.
3. Interpret the data on a plate reader.
4. Graph collected data and compute average doubling time for each yeast strain.
5. Repeat this process again and analyze the data.

Part 2

1. Finally we did the same process but in the 2 rows of wells surrounding our yeast we put 100ul of distilled water to try to avoid evaporation of our yeast to try to avoid the same error happening a third time