(r19) ProtocolsRecipesDavisMingioli < Lab

---+++ DM: Davis-Mingioli

Growth medium used by the long-term _E. coli_ evolution experiment.

|  *1L*  |  *5L*  |  *Component*  |  *MW*  |
|  5.3 g  |  26.7 g  | Potassium Phosphate (dibasic) K<sub>2</sub>HPO<sub>4</sub> | MW 174.18 |  
|  2 g  | 10 g | Potassium Phosphate (monobasic) KH<sub>2</sub>PO<sub>4</sub> | MW 136.07 |
|  1 g  | 5 g | Ammonium Sulfate (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>| MW 132.08 |
|  0.5 g  | 2.5 g | Sodium Citrate (trisodium, dihydrate) Na<sub>3</sub>C<sub>6</sub>H<sub>5</sub>O<sub>7</sub>  (H<sub>2</sub>O)<sub>2</sub>| MW 294.10 |
Add dH<sub>2</sub>O to final volume and autoclave. <br>

*Note:  if using Potassium phosphate (dibasic) trihydrate use 7g per L* 


After autoclaving add the following stock solutions: <br>

|  *1L*  |  *5L*  |  *Component*  |
| 1.0 ml | 5 ml | 10% (w/v) Magnesium Sulfate <nop>MgSO<sub>4</sub> (separately autoclaved stock) | 
| 1.0 ml | 5 ml | 0.2% (w/v) Thiamine (vitamin B1) (filter sterilized) |

And supplement with a carbon source.

If preparing DM-glucose (MW 180.16 g/mol), add this volume of *10% glucose* solution (separately autoclaved stock) to get the final concentration desired:

|  *1L*  |  *5L*  |  *DMX*  | *[glucose] (w/v)* | *[glucose] (mg/L)* | *[glucose] (M)* |
| 250µl | 1.25 ml | DM25 | 0.0025% | 25 mg/L | 139 µM |
| 1 ml | 5 ml | DM100 | 0.010% | 100 mg/L | 694 µM | 
| 2.5 ml | 12.5 ml | DM250 | 0.025% | 250 mg/L | 1.39 µM |
| 5 ml | 25 ml | DM500 | 0.05% | 500 mg/L | 2.78 mM |
| 10 ml | 50 ml | DM1000 | 0.1% | 1000 mg/L | 5.55 mM |
| 20 ml | 100 ml | DM2000 | 0.2% | 2000 mg/L | 11.1 mM |

Remember: DMX = DM + X mg/L glucose. Glucose may no longer limit the final growth density above approximately DM1000.

|  *Final composition*  ||
| 5.1 mM | Sodium (Na<sup>+</sup>) |
| 75.8 mM | Potassium (K<sup>+</sup>) |
| 15.2 mM | Ammonium (NH<sub>4</sub>) | 
| 0.83 mM | Magnesium (Mg<sup>2+</sup>) |
| 8.41 mM | Sulfate (SO<sub>4</sub><sup>2-</sup>) |
| 45.3 mM | Phosphate (PO<sub>4</sub><sup>3-</sup>) |
| 1.70 mM | Citrate |
| 139 µM | Glucose (in DM25) |

Barrick Lab > ProtocolsMediaRecipes > ProtocolsRecipesDavisMingioli

Topic revision: r19 - 2015-06-26 - 18:10:14 - Main.JeffreyBarrick

Lab.ProtocolsRecipesDavisMingioli moved from Lab.ProtocolsRecipesDavisMinimal on 2012-10-28 - 20:34 by Main.JeffreyBarrick -