Difference between revisions of "UW Stout/Sucrose Fermentation SP22"

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(Make Fermentation Solution)
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===Results===
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We found that the 4% total sucrose in concentration produced the most CO2 after 48 hours.

Revision as of 12:09, 3 May 2022

Introduction

The purpose of this protocol is to test if knockout yeast strains undergo fermentation, and if they do how much ethanol is produced. This will tell us if the gene that was knocked out plays a role in fermentation.

Materials

  • 1 ml Yeast Culture (Approximately 1 million cells)
    • You may need to dilute with sterile H2O to approximate the 1 million cells. Keep total volume at 1 ml.
  • PBS Buffer Solution
  • Yeast Growth Solution
    • 0.85g YNB, 2.5 Ammonium Sulfate, 0.32g CSM, 5ml 100x Leucine, 100x Histine, 100x Uracil, 435 ml DI H2O
  • 40% Sucrose Solution

Pilot Experiment Procedure

Set up yeast

  1. Collect 1 ml of yeast solution
  2. Centrifuge for 3 minutes at 1000x gravity
  3. Remove supernatant
  4. Add 1 ml of PBS buffer to yeast pellet

Make Fermentation Solution

  1. Mix the following ingredients for each concentration *See table 1
  2. Vortex solution for 30 seconds
  3. Store in 30°C incubator
  4. Sample solution (for ethanol production) in 24 and 48 hours.

Table 1

Concentration 1 Concentration 2 Concentration 3 Concentration 4
Yeast 1ml 1ml 1ml 1ml
Growth solution 8ml 8ml 8ml 8ml
40% Sucrose Solution 1ml 0.5ml 0.25ml 0.125ml
H20 0ml 0.5ml 0.75ml 0.875ml
Total Volume 10ml 10ml 10ml 10ml
Total sucrose concentration in solution 4% 2% 1% 0.5%

Results

We found that the 4% total sucrose in concentration produced the most CO2 after 48 hours.