UW-Stout/Caffeine SP22 - Revision history

Teagueb at 01:39, 4 May 2022

2022-05-04T01:39:23Z

Teagueb at 20:31, 3 May 2022

2022-05-03T20:31:05Z

Teagueb at 20:30, 3 May 2022

2022-05-03T20:30:20Z

Teagueb at 20:22, 3 May 2022

2022-05-03T20:22:06Z

Teagueb at 20:18, 3 May 2022

2022-05-03T20:18:11Z

Teagueb at 20:16, 3 May 2022

2022-05-03T20:16:35Z

Teagueb: Created page with "'''Materials:''' • Caffeine • Sterile H2O • Wild-Type Yeast Cells • Modified Yeast Cells '''Equipment:''' • Large Test Tube • 1.7 ml Microcentrifuge Tube • 200..."

2022-05-03T19:54:03Z

Created page with "'''Materials:''' • Caffeine • Sterile H2O • Wild-Type Yeast Cells • Modified Yeast Cells '''Equipment:''' • Large Test Tube • 1.7 ml Microcentrifuge Tube • 200..."

New page

'''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:'''
# Create a 32 mM Solution of Caffeine in Sterile H2O.
# Set Well Plate with 50 ul of Wild-Type Yeast Cells and the following concentrations of Caffeine.

{| border="1" cellspacing="4" cellpadding="2"
|
|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
|}

[[Image:Stress Test.png|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:'''
# Create a 4 mM Solution in Sterile H2O.
# Set Well Plates with 50 uL of each Yeast group of the following:
## BY4735
## YDL109C
## YGL140C
## YOR111W
## YHL029C
## YDR307W
## YNL058C
## YCL049C
## YGR079W
## YBL113C
# Add 50 uL of 4 mM Caffeine Solution to each Well.
# Incubate

@@ Line 45: / Line 45: @@
 # Create a 4 mM Solution in Sterile H2O.
 # Set Well Plates with 50 uL of each Yeast group of the following:
-## [[BY4735]]
+## BY4735
 ## [[YDL109C]]
 ## [[YGL140C]]

@@ Line 38: / Line 38: @@
 |}
-[[Image:Stress Test.png|Graph 1]]
+[[Image:Stresstest1.jpeg]]
 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.

@@ Line 45: / Line 45: @@
 # Create a 4 mM Solution in Sterile H2O.
 # Set Well Plates with 50 uL of each Yeast group of the following:
-## BY4735
+## [[BY4735]]
-## YDL109C
+## [[YDL109C]]
-## YGL140C
+## [[YGL140C]]
-## YOR111W
+## [[YOR111W]]
-## YHL029C
+## [[YHL029C]]
-## YDR307W
+## [[YDR307W]]
-## YNL058C
+## [[YNL058C]]
-## YCL049C
+## [[YCL049C]]
-## YGR079W
+## [[YGR079W]]
 ## [[YBL113C]]
 # Add 50 uL of 4 mM Caffeine Solution to each Well.
 # Incubate