Difference between revisions of "UW Stout/Ethanol FA19"
Line 45: | Line 45: | ||
|} | |} | ||
[[Image:BY4735.et.graph.png]] | [[Image:BY4735.et.graph.png]] | ||
+ | |||
This graph shows the ethanol tolerance in the Wild Type Yeast Cells (BY4735). | 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. | + | 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. We chose the 7% ethanol to use on the yeast knockout strands. |
==Knockout Strand Yeast Torture== | ==Knockout Strand Yeast Torture== | ||
===Protocol:=== | ===Protocol:=== |
Revision as of 15:43, 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. We chose the 7% ethanol to use on the 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 |