Difference: ProtocolsRecipesS2 (6 vs. 7)

Revision 72013-09-05 - GabrielSuarez

S2

Used for Acinetobacter

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1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O

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Recipe for 900mL

(Autoclave in three separate bottles - 300mL each)

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10 g

NH₄Cl

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After autoclaving, wait for containers to return to room temperature, then add the following sterile ingredients.

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Bottle 1 (Erlenmeyer flask: 300mL)

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

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1L	Component
1ml	10% MgSO₄
0.09ul	1M CaCl₂

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Bottle 2 (Erlenmeyer flask: 300mL)

300mL	Component
0.09g	MgSO₄

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0.5ml	0.1% FeSO₄ 7H₂O
5 ml	90% Lactic Acid

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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 (SO₄^2-)
106.1 mM	Phosphate (PO₄^3-)
50.0 mM	Lactic Acid

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-- AurkoDasgupta - 03 Jun 2013

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-- GabrielSuarez - 05 Sep 2013

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