50 mM Na2HPO4, 50 mM KH2PO4, 50 mM NH4Cl, 5 mM Na2SO4, 2 mM Mg2SO4, 0.5% glycerol, 0.05% glucose, 0.2% lactose, trace metals
1) Mix together in a 2 Liter Erlenmeyer flask :
898 ml ddH2O 2.68 g NH4Cl (CAN USE 15N-labeled AMMONIUM CHLORIDE) 0.71 g Na2SO4 5 g Glycerol 0.5 g Glucose 2 g Lactose 2) Autoclave using a Liquid cycle for 45 minutes.
3) Cool to room temperature, then add:
1 ml 2 M MgSO4 (0.22µm FILTER STERILIZED) 1 ml 1000X Trace Metals Mix (0.22µm FILTER STERILIZED) 50 ml 1 M Na2HPO4 (0.22µm FILTER STERILIZED or AUTOCLAVED) 50 ml 1 M KH2PO4 (0.22µm FILTER STERILIZED or AUTOCLAVED) 4) Add the appropriate antibiotic(s) and the media is ready to use.
Based on "Protein production by auto-induction in high-density shaking cultures", F. William Studier,
Protein Expression and Purfication, 41 (2005) 207-234
© 2007 Joseph T.E. Roland |