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Module 4: Yeast and the gene

Media prep for yeast culturing

     To make K+ liquid media for growing cultures

  • 1. Make 3M KCl solution (22.35gKCl / 100ml H2O).
  • 2. Autoclave the KCl solution.
  • 3. Add sterile KCl to 2x Complete APG to make it 100mM
    • -50ml of APG to 3.3ml of 3M KCl
  • 4. Dilute the solution 1:1 with dH2O
    • -50ml of APG/KCl solution w/50ml dH2O

     To make cultures for growth

  • 1. Add 1.5 ml growth media in each 2 ml tube
  • 2. Pull a culture from each of the tubes to place in the tube
  • 3. Incubate at 30°C overnight (shaking)

     To make the low K+ agar plates

  • 1. Make a 50ml sterile 2% agar solution (autoclave)
  • 2. Add 3.3ml (100mM) or 1.15ml (50mM) 3M KCl solution.
  • 3. Add 50ml APG media.
  • 4. Pour plates (need 6 x 50mM and 6 x 100mM per class)

     To make the normal K+ plates

  • 1. Make a 50ml sterile 2% agar solution.
  • 2. Add 5g YPD media to 100ml of dH2O
  • 3. Add 50ml of the YPD solution to the agar.
  • 4. Autoclave the mixture.
  • 5. Add KCl to get 150mM.
  • 6. Pour plates (need 6 per class)