Difference between revisions of "UW-Stout/Caffeine SP22"

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## YCL049C
 
## YCL049C
 
## YGR079W
 
## YGR079W
## YBL113C
+
## [[YBL113C]]
 
# Add 50 uL of 4 mM Caffeine Solution to each Well.
 
# Add 50 uL of 4 mM Caffeine Solution to each Well.
 
# Incubate
 
# Incubate

Revision as of 13:22, 3 May 2022

Materials: • Caffeine • Sterile H2O • Wild-Type Yeast Cells • Modified Yeast Cells

Equipment: • Large Test Tube • 1.7 ml Microcentrifuge Tube • 200 ul Pipette • Micro Pipette Tips • Well Plates

Calibration Protocol:

  1. Create a 32 mM Solution of Caffeine in Sterile H2O.
  2. Set Well Plate with 50 ul of Wild-Type Yeast Cells and the following concentrations of Caffeine.
Well 1 Well 2 Well 3 Well 4 Well 5 Well 6 Well 7 Well 8
Caffeine Concentration 0 mM 4 mM 8 mM 12 mM 16 mM 20 mM 28 mM 32 mM

Graph 1

As can be seen in the graph above, Well 1 (our control group) shows the expected growth curve. Every group with a concentration of at least 8mM showed little growth and can be assumed dead. Well 2, at 4mM, shows a reduction in growth but not a visible result for future tests.

Knockout Protocol:

  1. Create a 4 mM Solution in Sterile H2O.
  2. Set Well Plates with 50 uL of each Yeast group of the following:
    1. BY4735
    2. YDL109C
    3. YGL140C
    4. YOR111W
    5. YHL029C
    6. YDR307W
    7. YNL058C
    8. YCL049C
    9. YGR079W
    10. YBL113C
  3. Add 50 uL of 4 mM Caffeine Solution to each Well.
  4. Incubate