Difference: ProtocolsRecipesDavisMingioli (20 vs. 21)

Revision 212024-09-10 - SophiaLin

META TOPICPARENT	name="ProtocolsMediaRecipes"

DM: Davis-Mingioli

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

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0.5L	1L	5L	Component	MW
2.67 g	5.34 g	26.7 g	Potassium Phosphate (dibasic) K₂HPO₄	MW 174.18
1 g	2 g	10 g	Potassium Phosphate (monobasic) KH₂PO₄	MW 136.07
0.5 g	1 g	5 g	Ammonium Sulfate (NH₄)₂ SO₄	MW 132.08
0.25 g	0.5 g	2.5 g	Sodium Citrate (trisodium, dihydrate) Na₃C₆H₅O₇ (H₂O)₂	MW 294.10

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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

Add dH₂O to final volume and autoclave.

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Note: if using Potassium phosphate (dibasic) trihydrate (MW 228.22) use 7g per L

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Note: 4.4 L of DM makes 8 bottles with 550 ml each.

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Note: If using Potassium phosphate (dibasic) trihydrate (MW 228.22) use 7g per L

Deleted:

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After autoclaving add the following stock solutions:

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0.5L	1L	5L	Component

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After autoclaving add the following stock solutions:

Deleted:

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0.5 ml	1.0 ml	5 ml	10% (w/v) Magnesium Sulfate MgSO₄ (separately autoclaved stock)
0.5 ml	1.0 ml	5 ml	0.2% (w/v) Thiamine (vitamin B1) (filter sterilized)

Added:

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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:

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0.5L	1L	5L	DMX	[glucose] (w/v)	[glucose] (mg/L)	[glucose] (M)
125 µl	250 µl	1.25 ml	DM25	0.0025%	25 mg/L	139 µM
0.5 ml	1 ml	5 ml	DM100	0.010%	100 mg/L	694 µM
1.25 ml	2.5 ml	12.5 ml	DM250	0.025%	250 mg/L	1.39 µM
2.5 ml	5 ml	25 ml	DM500	0.05%	500 mg/L	2.78 mM
5 ml	10 ml	50 ml	DM1000	0.1%	1000 mg/L	5.55 mM
10 ml	20 ml	100 ml	DM2000	0.2%	2000 mg/L	11.1 mM

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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 (SO₄^2-)
45.3 mM	Phosphate (PO₄^3-)
1.70 mM	Citrate
139 µM	Glucose (in DM25)

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

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