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| META TOPICPARENT |
name="ProtocolsMediaRecipes" |
DM: Davis Minimal |
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>
> | Notes: Salts, agar and sugars need to be prepared and autoclaved separately.
For making 1.5 liters of DM agar do the following:
Combine the following in a 2 L flask (including magnetic stir bar):
| 10.5 g |
Potassium Phosphate (dibasic, trihydrate) HK2PO4 (H2O)3 |
MW 228.17 |
| 3 g |
Potassium Phosphate (monobasic) H2KPO4 |
MW 136.07 |
| 1.5 g |
Ammonium Sulfate (NH4)2 SO4 |
MW 132.08 |
| 0.75 g |
Sodium Citrate (trisodium, dihydrate) Na3C6H5O7 (H2O)2 |
MW 294.10 |
Add 500 ml of dH2O, cover with tin foil and autoclave.
Next, prepare the agar in a 1 liter flask (including magnetic stir bar) by adding the following:
| 24 g |
Agar |
| 1.5 ml |
Antifoam (5%) |
Add 500 ml of dH2O, cover with tin foil and autoclave.
Next, prepare the sugar in a separate 1 liter flask by adding the following:
| 6 g |
Glucose (or other sugar) |
Add 500 ml of dH2O, cover with tin foil and autoclave.
After autoclaving combine the contents of the three flasks into the larger flask and add the following stock solutions:
| 1.5 ml |
10% Magnesium Sulfate MgSO4 |
MW 120.37 |
|
| 1.5 ml |
0.2% Thiamin (vitamin B1) |
Note* Thiamin must be syringe filtered into the media |
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To make 1 liter of DM plates combine the following in a 2 L flask (including magnetic stir bar):
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| 7 g |
Potassium Phosphate (dibasic, trihydrate) HK2PO4 (H2O)3 |
MW 228.17 |
| 2 g |
Potassium Phosphate (monobasic) H2KPO4 |
MW 136.07 |
| 1 g |
Ammonium Sulfate (NH4)2 SO4 |
MW 132.08 |
| 0.5 g |
Sodium Citrate (trisodium, dihydrate) Na3C6H5O7 (H2O)2 |
MW 294.10 |
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<
< | Add dH2O to 1 liter. Autoclave. Then add
| 1 ml |
10% Magnesium Sulfate MgSO4 |
MW 120.37 |
Note* The MgSO4 must be autoclaved separately. |
| 1 ml |
0.2% Thiamin (vitamin B1) |
|
|
>
> | Add about 333 ml of dH2O, cover with tin foil and autoclave.
Next, prepare the agar in a 1 liter flask (including magnetic stir bar) by adding the following:
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|
>
> |
| 16 g |
Agar |
| 1 ml |
Antifoam (5%) |
Add about 333 ml of dH2O, cover with tin foil and autoclave.
Next, prepare the sugar in a separate 1 liter flask by adding the following:
| 4 g |
Glucose (or other sugar) |
Add about 333 ml of dH2O, cover with tin foil and autoclave.
After autoclaving combine the contents of the three flasks into the larger flask and add the following stock solutions:
| 1 ml |
10% Magnesium Sulfate MgSO4 |
MW 120.37 |
|
| 1 ml |
0.2% Thiamin (vitamin B1) |
Note* Thiamin must be syringe filtered into the media |
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Final nutrient concentrations:
| 67.4 mM |
Phosphate (PO43-) |
| 8.38 mM |
Sulfate (SO42-) |
| 15.1 mM |
Ammonium (NH4) |
| 5.1 mM |
Sodium (Na+) |
| 98.1 mM |
Potassium (K+) |
| 0.83 mM |
Magnesium (MGgsup>2+) |
| 1.7 mM |
Citrate |
| 139 µM |
Glucose (in DM25) |
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