# UW Stout/DMSO FA19

## Preliminary Yeast Torture Protocol

**Materials:**

- Wild Type Yeast
- DMSO at 100% solution
- Sterile H
_{2}O - 96 well plate

**Equipment**

- P2 Micropipette
- P10 Micropipette
- P200 Micropipette
- Molecular Devices SpectraMax Plus 384 Microplate Reader

**Protocol**

- Label wells in accordance to percent DMSO dilution
- Add 50 μliters of wild type yeast into the wells on the 96 well plate
- Add 49 μliters of sterile H
_{2}O into well labelled for 1% DMSO - Add 1 μliters of DMSO at 100% into well labelled 1% DMSO
- Follow steps 2-4 for 0%, 0.1%, 0.2%, 0.5%, 2%, 5%, 10%, 20%, 50% DMSO desired
- Incubate in microplate reader for 6-12 hours at 30°C
- Collect and analyze data to determine optimal DMSO% for stressing yeast

**Calculations**

- C
_{1}V_{1}=C_{2}V_{2}

- Make 50% solution of DMSO
- Make 10% solution of DMSO

- 100 μliters=V
_{2}

- 0%= 50 μliters of water, and 50 μliters of yeast solution
- 0.1% = 49 μliters of water, 1 μliters of DMSO at 10%, and 50 μliters of yeast solution
- 0.2%= 48 μliters of water, 2 μliters of DMSO at 10%, and 50 μliters of yeast solution
- 0.5%=49 μliters of water, 1 μliters of DMSO at 50%, and 50 μliters of yeast solution
- 1%= 49 μliters of water, 1 μliter of DMSO at 100%, 50 μliters of yeast solution
- 2%= 48 μliters of water, 2 μliters of DMSO at 100%, and 50 μliters of yeast solution
- 5%= 45 μliters of water, 5 μliters of DMSO at 100%, and 50 μliters of yeast solution
- 10% = 40 μliters of water, 10 μliters of DMSO at 100%, 50 μliters of yeast solution
- 20%= 30 μliters of water, 20 μliters of DMSO at 100%, 50 μliters of yeast solution
- 50%= 50 μliters of DMSO at 100%, and 50 μliters of yeast solution