Wittrup: Yeast Minimal Media

From OpenWetWare
Jump to: navigation, search


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)