Difference: ProtocolsRecipesDavisMingioli (10 vs. 11)

Revision 112012-11-20 - JeffreyBarrick

META TOPICPARENT	name="ProtocolsMediaRecipes"

DM: Davis-Mingioli

Growth medium used by the long-term E. coli evolution experiment.

1L	5L	Component	MW
7 g	35 g	Potassium Phosphate (dibasic, trihydrate) HK₂PO₄ (H₂O)₃	MW 228.17
2 g	10 g	Potassium Phosphate (monobasic) H₂KPO₄	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.

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)

Changed:

<
<

25 µl	125 µl	DM25	0.0025%	25 mg/L	139 µM
100 µl	500 µl	DM100	0.010%	100 mg/L	694 µM
250 µl	1.25 ml	DM250	0.025%	250 mg/L	1.39 µM
500 µl	2.5 ml	DM500	0.05%	500 mg/L	2.78 mM
1.0 ml	5 ml	DM1000	0.1%	1000 mg/L	5.55 mM
2.0 ml	10 ml	DM2000	0.2%	2000 mg/L	11.1 mM

>
>

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
67.4 mM	Phosphate (PO₄^3-)
8.38 mM	Sulfate (SO₄^2-)
15.1 mM	Ammonium (NH₄)
5.1 mM	Sodium (Na⁺)
98.1 mM	Potassium (K⁺)
0.83 mM	Magnesium (Mg²⁺)
1.7 mM	Citrate
139 µM	Glucose (in DM25)

META TOPICMOVED	by="JeffreyBarrick" date="1351456485" from="Lab.ProtocolsRecipesDavisMinimal" to="Lab.ProtocolsRecipesDavisMingioli"

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