N-5052 Autoinduction Media - 1 Liter 900 ml 800 ml 700 ml 600 ml 500 ml 400 ml 300 ml 200 ml 100 ml 50 ml 1000 ml

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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

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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