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) 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
Add dH2O to final volume and autoclave.

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)
250Ál 1.25 ml DM25 0.0025% 25 mg/L 139 ÁM
1 ml 5 ml DM100 0.010% 100 mg/L 694 ÁM
2.5 ml 12.5 ml DM250 0.025% 250 mg/L 1.39 ÁM
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-)
1.70 mM Citrate
139 ÁM Glucose (in DM25)

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Topic revision: r19 - 26 Jun 2015 - 18:10:14 - Main.JeffreyBarrick
Lab.ProtocolsRecipesDavisMingioli moved from Lab.ProtocolsRecipesDavisMinimal on 28 Oct 2012 - 20:34 by Main.JeffreyBarrick - put it back
 
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