UW-Stout/Isopropanol SP23

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%.

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

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

1. In the third microcentrifuge tube add 20ul of 100% isopropyl to 80ul of sterile water.

1. In the fourth microcentrifuge tube add 10ul of 100% isopropyl to 90ul of sterile water

1. In the fifth microcentrifuge tube add 4ul of 100% isopropyl to 96ul of sterile water

1. In the sixth microcentrifuge tube add 2ul of 100% isopropyl to 98ul of sterile water

1. In the seventh microcentrifuge tube add 2ul of 50% isopropyl to 98ul of sterile water

1. In the eighth microcentrifuge tube add 4ul of 10% isopropyl to 96ul of sterile water

1. In the ninth microcentrifuge tube add 2ul of 10% isopropyl to 98ul of sterile water

1. In the tenth microcentrifuge tube add 4ul of 5% isopropyl to 96ul of sterile water

1. In the eleventh microcentrifuge tube add 2ul of 5% isopropyl to 98ul of sterile water

1. 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