Growth medium used by the long-term E. coli evolution experiment.
1L | 5L | Component | MW |
---|---|---|---|
5.3 g | 26.7 g | Potassium Phosphate (dibasic) K2HPO4 | MW 174.18 |
2 g | 10 g | Potassium Phosphate (monobasic) KH2PO4 | MW 136.07 |
1 g | 5 g | Ammonium Sulfate (NH4)2 SO4 | MW 132.08 |
0.5 g | 2.5 g | Sodium Citrate (trisodium, dihydrate) Na3C6H5O7 (H2O)2 | MW 294.10 |
Note: if using Potassium phosphate (dibasic) trihydrate use 7g per L
After autoclaving add the following stock solutions:
1L | 5L | Component |
---|---|---|
1.0 ml | 5 ml | 10% (w/v) Magnesium Sulfate MgSO4 (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+) |
75.8 mM | Potassium (K+) |
15.2 mM | Ammonium (NH4) |
0.83 mM | Magnesium (Mg2+) |
8.41 mM | Sulfate (SO42-) |
45.3 mM | Phosphate (PO43-) |
1.7 mM | Citrate |
139 µM | Glucose (in DM25) |
Barrick Lab > ProtocolsMediaRecipes > ProtocolsRecipesDavisMingioli