Difference between revisions of "UW Stout/Ethanol FA19"
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| Line 3: | Line 3: | ||
Materials: | Materials: | ||
# neat ethanol | # neat ethanol | ||
| − | # yeast | + | # yeast (WT:BY4735) |
# sterile water | # sterile water | ||
# 200 ul | # 200 ul | ||
| Line 44: | Line 44: | ||
| 50 | | 50 | ||
|} | |} | ||
| + | [[Image:BY4735.et.graph.png]] | ||
| + | |||
| + | This graph shows the ethanol tolerance in the Wild Type Yeast Cells (BY4735). | ||
| + | This graph helped us determine the amount of yeast to add to our knockout strains. We determined this by examining the graph and found a percentage of ethanol that shows a change in growth and doesn't kill the cells. For this we chose the 7% ethanol to use on our yeast knockout strands. | ||
==Knockout Strand Yeast Torture== | ==Knockout Strand Yeast Torture== | ||
===Protocol:=== | ===Protocol:=== | ||
Materials: | Materials: | ||
| − | + | #Yeast Knockout Strains: | |
##YAL061W | ##YAL061W | ||
##YBL086C | ##YBL086C | ||
| Line 59: | Line 63: | ||
##YKR005C | ##YKR005C | ||
##WT:BY4735 | ##WT:BY4735 | ||
| − | + | #neat ethanol | |
| − | + | #sterile water | |
| − | + | #micropipeter(s) | |
| − | + | #96 well plate | |
Procedure: | Procedure: | ||
#add 50 ul of Knockout Strain to individual well (96 well plate). | #add 50 ul of Knockout Strain to individual well (96 well plate). | ||
Latest revision as of 15:45, 10 December 2019
Yeast torture 1
Protocol:
Materials:
- neat ethanol
- yeast (WT:BY4735)
- sterile water
- 200 ul
- 96 well plate
Procedure:
- (add 50ul mixture to 50ul yeast in 96 well plate.)x2. Used 12 wells in total
- fill 6 200ul tubes, each with water and ethanol concentration x2. Resulting in 6 200ul tubes with 100ul of mixture in each.
| 1 | 2 | 3 | 4 | 5 | 6 | |
|---|---|---|---|---|---|---|
| ul ethanol | 0 | 3.75 | 7 | 10.25 | 13.5 | 16.75 |
| ul water | 50 | 46.25 | 43 | 39.75 | 37.5 | 33.25 |
| ul yeast | 50 | 50 | 50 | 50 | 50 | 50 |
This graph shows the ethanol tolerance in the Wild Type Yeast Cells (BY4735). This graph helped us determine the amount of yeast to add to our knockout strains. We determined this by examining the graph and found a percentage of ethanol that shows a change in growth and doesn't kill the cells. For this we chose the 7% ethanol to use on our yeast knockout strands.
Knockout Strand Yeast Torture
Protocol:
Materials:
- Yeast Knockout Strains:
- YAL061W
- YBL086C
- YBL9081W
- YBR033W
- YBR138C
- YBR220C
- YHR048W
- YGL140C
- YJR061W
- YKR005C
- WT:BY4735
- neat ethanol
- sterile water
- micropipeter(s)
- 96 well plate
Procedure:
- add 50 ul of Knockout Strain to individual well (96 well plate).
- add 50 ul of water+ethanol mixture to individual well (Follow table below).
| Well | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| ul yeast | 50 | 50 | 50 | 50 | 50 | 50 | 50 | 50 | 50 | 50 | 50 | 50 |
| ul ethanol | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 |
| ul water | 43 | 43 | 43 | 43 | 43 | 43 | 43 | 43 | 43 | 43 | 43 | 43 |
