Growth medium used by the long-term E. coli evolution experiment.
0.5L | 1L | 4.4L | 5L | Component | MW |
---|---|---|---|---|---|
2.67 g | 5.34 g | 23.32 g | 26.7 g | Potassium Phosphate (dibasic) K2HPO4 | MW 174.18 |
1 g | 2 g | 8 g | 10 g | Potassium Phosphate (monobasic) KH2PO4 | MW 136.07 |
0.5 g | 1 g | 4.4 g | 5 g | Ammonium Sulfate (NH4)2 SO4 | MW 132.08 |
0.25 g | 0.5 g | 2.2 g | 2.5 g | Sodium Citrate (trisodium, dihydrate) Na3C6H5O7 (H2O)2 | MW 294.10 |
Note: 4.4 L of DM makes 8 bottles with 550 ml each.
Note: If using Potassium phosphate (dibasic) trihydrate (MW 228.22) use 7g per L
After autoclaving add the following stock solutions:
0.5L | 550mL | 1L | 5L | Component |
---|---|---|---|---|
0.5 ml | 550 µl | 1.0 ml | 5 ml | 10% (w/v) Magnesium Sulfate MgSO4 (separately autoclaved stock) |
0.5 ml | 1000 µl | 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:
0.5L | 550mL | 1L | 5L | DMX | [glucose] (w/v) | [glucose] (mg/L) | [glucose] (M) |
---|---|---|---|---|---|---|---|
125 µl | 137.5 µl | 250 µl | 1.25 ml | DM25 | 0.0025% | 25 mg/L | 139 µM |
0.5 ml | 0.55 ml | 1 ml | 5 ml | DM100 | 0.010% | 100 mg/L | 694 µM |
1.25 ml | 1.375 ml | 2.5 ml | 12.5 ml | DM250 | 0.025% | 250 mg/L | 1.39 µM |
2.5 ml | 2.75 ml | 5 ml | 25 ml | DM500 | 0.05% | 500 mg/L | 2.78 mM |
5 ml | 5.5 ml | 10 ml | 50 ml | DM1000 | 0.1% | 1000 mg/L | 5.55 mM |
10 ml | 11 ml | 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-) |
1.70 mM | Citrate |
139 µM | Glucose (in DM25) |