UW-Stout/Copper FA21

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Revision as of 14:33, 21 December 2021 by Teagueb (talk | contribs) (Interpreting Gene Removed Data)
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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.

Interpreting Wild Type Data

Wild.jpg

As we can see in the graph we are looking to see where the yeast cell was affected by the copper but not outright kill it. In the graph we can see at the 0.5mM concentration that we have growth in the form of an S-curve and then it starts to stop growing which means that the yeast was affect meaning that this was our optimal point for yeast and copper sulfate.

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 YFG1.jpg

In the above graph, you can see that there is an anomaly in the data that at first we assumed was either a pipetting or instrument error. However, when running the experiment again we got the same result and we believe the growth is drying up.
  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

Interpreting Gene Removed Data

YHL012W-2.jpgYHL012W-normal.jpg