Used for Acinetobacter

Recipe for 900mL

(Autoclave in three separate bottles - 300mL each)

Bottle 1 (Erlenmeyer flask: 300mL)

300mL Component
1.35g KH2PO4 (monobasic)
22.95g Na2HPO4 7H2O

Bottle 2 (Erlenmeyer flask: 300mL)

300mL Component
0.09g MgSO4

Bottle 3 (1L bottle: 300mL)

300mL Component
1.8g NH4Cl
81ul 1M CaCl2

After autoclaving, let these reach room temperature and transfer all into one 1L bottle (Bottle 3), then, finally add the following sterile components:

0.45ml 0.1% FeSO4 7H2O
4.5 ml 90% Lactic Acid

The functional constraints are that Calcium Chloride cannot be autoclaved in the presence of sulfates, or it will precipitate as calcium sulfate. Ferrous sulfate when autoclaved will decompose to ferric sulfate, ferric oxide and sulfur dioxide. This means that either calcium chloride OR magnesium sulfate may be added before autoclaving, but not both.

Final composition
190.2 mM Sodium (Na+)
11.0 mM Potassium (K+)
37.8 mM Ammonium (NH4+)
0.09 mM Calcium (Ca2+)
0.83 mM Magnesium (Mg2+)
0.033 mM Iron II (Fe2+)
38.0 mM Chloride (Cl-)
0.87 mM Sulfate (SO42-)
106.1 mM Phosphate (PO43-)
50.0 mM Lactic Acid

-- Main.GabrielSuarez - 05 Sep 2013

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