Barrick Lab :: ProtocolsRecipesS2

<noautolink>
---+++ S2

Used for Acinetobacter

Recipe for 900mL
 
(Autoclave in three separate bottles - 300mL each) 

*Bottle 1* (Erlenmeyer flask: 300mL)
|  *300mL*  |  *Component*  |
| 1.35g | KH<sub>2</sub>PO<sub>4</sub> (monobasic) | 
| 22.95g | Na<sub>2</sub>HPO<sub>4</sub> 7H<sub>2</sub>O| 

*Bottle 2* (Erlenmeyer flask: 300mL)
|  *300mL*  |  *Component*  |
| 0.09g | MgSO<sub>4</sub> | 

*Bottle 3* (1L bottle: 300mL)
|  *300mL*  |  *Component*  |
| 1.8g | NH<sub>4</sub>Cl| 
| 81ul | 1M CaCl<sub>2</sub> | 

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% FeSO<sub>4</sub> 7H<sub>2</sub>O |
| 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<sup>+</sup>) |
| 11.0 mM | Potassium (K<sup>+</sup>) |
| 37.8 mM | Ammonium (NH<sub>4</sub><sup>+</sup>) | 
| 0.09 mM | Calcium (Ca<sup>2+</sup>) |
| 0.83 mM | Magnesium (Mg<sup>2+</sup>) |
| 0.033 mM | Iron II (Fe<sup>2+</sup>) |
| 38.0 mM | Chloride (Cl<sup>-</sup>)|
| 0.87 mM | Sulfate (SO<sub>4</sub><sup>2-</sup>) |
| 106.1 mM | Phosphate (PO<sub>4</sub><sup>3-</sup>) |
| 50.0 mM | Lactic Acid|

-- Main.GabrielSuarez - 05 Sep 2013

Barrick Lab > ProtocolsRecipesS2

Contributors to this topic

AurkoDasgupta, GabrielSuarez

Topic revision: r7 - 2013-09-05 - 22:31:47 - Main.GabrielSuarez