Difference between revisions of "UW-Stout/Isopropanol SP23"
| Line 1: | Line 1: | ||
| − | + | Introduction | |
We are going to introduce the wild type yeast to isopropyl alcohol as a stress to see how it reacts if at all. | We are going to introduce the wild type yeast to isopropyl alcohol as a stress to see how it reacts if at all. | ||
| − | + | Materials | |
*100% Isopropyl alcohol | *100% Isopropyl alcohol | ||
| Line 18: | Line 18: | ||
*12 microcentrifuge tubes | *12 microcentrifuge tubes | ||
| − | + | Procedure | |
| − | + | Diluting the Isopropyl alcohol | |
*Compute the volume of your 100% isopropyl to achieve a dilution of 50%, 10%, and 5%. | *Compute the volume of your 100% isopropyl to achieve a dilution of 50%, 10%, and 5%. | ||
| − | + | *In the first microcentrifuge tube add 100ul of 100% isopropyl. | |
| − | + | *In the second microcentrifuge tube add 40ul of 100% isopropyl to 60ul of sterile water. | |
| − | + | *In the third microcentrifuge tube add 20ul of 100% isopropyl to 80ul of sterile water. | |
| − | + | *In the fourth microcentrifuge tube add 10ul of 100% isopropyl to 90ul of sterile water | |
| − | + | *In the fifth microcentrifuge tube add 4ul of 100% isopropyl to 96ul of sterile water | |
| − | + | *In the sixth microcentrifuge tube add 2ul of 100% isopropyl to 98ul of sterile water | |
| − | + | *In the seventh microcentrifuge tube add 2ul of 50% isopropyl to 98ul of sterile water | |
| − | + | *In the eighth microcentrifuge tube add 4ul of 10% isopropyl to 96ul of sterile water | |
| − | + | *In the ninth microcentrifuge tube add 2ul of 10% isopropyl to 98ul of sterile water | |
| − | + | *In the tenth microcentrifuge tube add 4ul of 5% isopropyl to 96ul of sterile water | |
| − | + | *In the eleventh microcentrifuge tube add 2ul of 5% isopropyl to 98ul of sterile water | |
| − | + | *In the twelfth microcentrifuge tube add 100ul of sterile water for the control | |
| − | + | Mixing diluted isopropyl and yeast cells | |
Add 50ul of wild type yeast to each well labeled 1-12 | Add 50ul of wild type yeast to each well labeled 1-12 | ||
Revision as of 11:04, 25 April 2023
Introduction We are going to introduce the wild type yeast to isopropyl alcohol as a stress to see how it reacts if at all.
Materials
- 100% Isopropyl alcohol
- Room temp
- Wild Type yeast tube
- BY4735
- Sterile water
- 96 well plate
- 12 microcentrifuge tubes
Procedure
Diluting the Isopropyl alcohol
- Compute the volume of your 100% isopropyl to achieve a dilution of 50%, 10%, and 5%.
- In the first microcentrifuge tube add 100ul of 100% isopropyl.
- In the second microcentrifuge tube add 40ul of 100% isopropyl to 60ul of sterile water.
- In the third microcentrifuge tube add 20ul of 100% isopropyl to 80ul of sterile water.
- In the fourth microcentrifuge tube add 10ul of 100% isopropyl to 90ul of sterile water
- In the fifth microcentrifuge tube add 4ul of 100% isopropyl to 96ul of sterile water
- In the sixth microcentrifuge tube add 2ul of 100% isopropyl to 98ul of sterile water
- In the seventh microcentrifuge tube add 2ul of 50% isopropyl to 98ul of sterile water
- In the eighth microcentrifuge tube add 4ul of 10% isopropyl to 96ul of sterile water
- In the ninth microcentrifuge tube add 2ul of 10% isopropyl to 98ul of sterile water
- In the tenth microcentrifuge tube add 4ul of 5% isopropyl to 96ul of sterile water
- In the eleventh microcentrifuge tube add 2ul of 5% isopropyl to 98ul of sterile water
- In the twelfth microcentrifuge tube add 100ul of sterile water for the control
Mixing diluted isopropyl and yeast cells
Add 50ul of wild type yeast to each well labeled 1-12
Starting in well 1 add 50ul of the contents of microcentrifuge tube 1
Repeat this process with microcentrifuge tubes 2-12 adding 50ul in their respected wells labeled 2-12
