Wittrup: Yeast Minimal Media

SD-CAA, SG-CAA, SD-SCAA, SG-SCAA

Liquid, 1000 mL

• Carbon source
  • SD: 20g Dextrose (D-glucose)
  • SG: 20g Galactose
• Nitrogen source
  • 6.7g Yeast N2 base
• Amino acid source
  • 5.0g CAA (-ADE, -URA, -TRP)
  • Or, Synthetic CAA..
• Buffer
  • pH 6.0 phosphate buffer (standard)
    • 10.19g Na2HPO4.7H2O (which equals 5.40g Na2HPO4 anhydrous)
    • 8.56g NaH2PO4.H2O
  • pH 4.5 citrate buffer (optional: lower pH discourages bacterial growth)
    • 10.4g sodium citrate
    • 7.4g citric acid monohydrate

Add ddH2O to final volume. Filter sterilize.

Plates, 1000 mL

• Filter Sterilize:
  • Carbon source, Nitrogen source, Amino acid source in 0.1*(Final Volume)
• Autoclave:
  • Buffer salts, 15-18g agar, 182g sorbitol in 0.9*(Final Volume)
• Add first mixture to the autoclaved mixture after it has cooled to below 50 oC (touchable by bare hand)