Difference between revisions of "EMM - Edinburgh Minimal Medium"
| Line 15: | Line 15: | ||
|Na<sub>2</sub>HPO<sub>4</sub>.2H<sub>2</sub>O (or anhydrous)||2.76 g (2.2 g) | |Na<sub>2</sub>HPO<sub>4</sub>.2H<sub>2</sub>O (or anhydrous)||2.76 g (2.2 g) | ||
|- | |- | ||
| − | | | + | |NH<sub>4</sub>Cl || 5.0 g |
|- | |- | ||
|[[Salts Stock x50]] ||20 ml | |[[Salts Stock x50]] ||20 ml | ||
| Line 22: | Line 22: | ||
|- | |- | ||
|[[Mineral Stock x10.000]] ||0.1 ml | |[[Mineral Stock x10.000]] ||0.1 ml | ||
| − | |||
| − | |||
|} | |} | ||
| − | Sterilize by autoclaving for 15min at 121C.<BR> | + | Sterilize by autoclaving for 15min at 121C. |
| − | Add | + | |
| − | <BR> | + | <BR>Add 26.6 ml sterile [[Glucose Stock Solution 60%|60% Glucose]] after autoclaving the medium (if added before, this interacts with ammonium chloride resulting in almost black medium).<BR> |
| + | |||
| + | Add supplements as required (225mg/L for most, 100mg/L for Adenine) before autoclaving.<BR> | ||
| − | |||
For solid medium add 2% Agar-agar (20g/L) | For solid medium add 2% Agar-agar (20g/L) | ||
Revision as of 14:54, 13 September 2022
Edinburgh Minimal Medium (EMM, EMM2, or EMM Phthallate)
This is a synthetic defined minimal medium EMM2 is used for vegetative growth.
The medium is also used to screen strains for deficiencies and prototrophic markers.
We use this medium during the asexual phase of all our experimental evolution.
Furthermore, Edinburgh Minimal Media is a thiamine-free medium that does not suppress the nmt1 promoter.
The medium can be modified to a Mating medium by leaving out ammoniumchloride as a nitrogen source (EMM-N) or modified to PMG.
| Ingredient | Amount per liter |
|---|---|
| Potassium Hydrogen Phthallate | 3.0 g |
| Na2HPO4.2H2O (or anhydrous) | 2.76 g (2.2 g) |
| NH4Cl | 5.0 g |
| Salts Stock x50 | 20 ml |
| Vitamin Stock x1000 | 1.0 ml |
| Mineral Stock x10.000 | 0.1 ml |
Sterilize by autoclaving for 15min at 121C.
Add 26.6 ml sterile 60% Glucose after autoclaving the medium (if added before, this interacts with ammonium chloride resulting in almost black medium).
Add supplements as required (225mg/L for most, 100mg/L for Adenine) before autoclaving.
For solid medium add 2% Agar-agar (20g/L)
