UW-Stout/Isopropanol SP23

Wild Type Yeast Isopropanol Stress Experiment

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%.

Creating Stock Solutions

• In the first microcentrifuge tube add 100ul of 100% isopropyl.

• 2In the second microcentrifuge tube add 40ul of 100% isopropyl to 60ul of sterile water.

• 3In 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