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| META TOPICPARENT |
name="ProtocolsMediaRecipes" |
MG: Minimal Glucose agar, aka DM: Davis Mingioli agar
Note: When making these plates it is necessary to prepare and autoclave the 3 main parts (salts, agar, and sugar) separately. Compounds that inhibit growth are produced when agar and phosphate or phosphate and glucose are autoclaved together. |
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<
< |
| 500 ml |
1L |
1.5L |
Component |
MW |
| 2.65 g |
5.3 g |
8 g |
Potassium Phosphate (dibasic) K2HPO4 |
MW 174.18 |
| 1 g |
2 g |
3 g |
Potassium Phosphate (monobasic) KH2PO4 |
MW 136.07 |
| 0.5 g |
1 g |
1.5 g |
Ammonium Sulfate (NH4)2 SO4 |
MW 132.08 |
| 0.25 g |
0.5 g |
0.75 g |
Sodium Citrate (trisodium, dihydrate) Na3C6H5O7 (H2O)2 |
MW 294.10 |
| 166.7 ml |
333 ml |
500 ml |
dH2O |
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|
>
> |
| 500 mL |
1.0L |
Component |
| 2.65 g |
5.3 g |
Potassium Phosphate (dibasic) K2HPO4 |
| 1 g |
2 g |
Potassium Phosphate (monobasic) KH2PO4 |
| 0.5 g |
1 g |
Ammonium Sulfate (NH4)2 SO4 |
| 0.25 g |
0.5 g |
Sodium Citrate (trisodium, dihydrate) Na3C6H5O7 (H2O)2 |
| 166.7 mL |
333 mL |
dH2O |
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Separately, prepare the agar:
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<
< |
| 500 ml |
1L |
1.5L |
Component |
| 8 g |
16 g |
24 g |
Agar |
| 500 µl |
1 ml |
1.5 ml |
Antifoam (5%) |
| 166.7 ml |
333 ml |
500 ml |
dH2O |
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>
> |
| 500 mL |
1.0 L |
Component |
| 8 g |
16 g |
Agar |
| 500 µL |
1 mL |
Antifoam (5%) |
| 166.7 mL |
333 mL |
dH2O |
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|
<
< | Autoclave.
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Next, prepare the sugar solution. The final concentration is 0.4% w/v:
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|
<
< |
| 500 ml |
1L |
1.5L |
Component |
| 2 g |
4 g |
6 g |
Glucose, arabinose, or other sugar |
| 166.7 ml |
333 ml |
500 ml |
dH2O |
Autoclave.
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>
> |
| 500 mL |
1.0 L |
Component |
| 2 g |
4 g |
Glucose, arabinose, or other sugar |
| 166.7 mL |
333 mL |
dH2O |
Autoclave the 3 parts separately for a 40 minute sterilization time.
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After the 3 parts have been autoclaved, combine the contents of the three flasks together while they are still warm add the following stock solutions:
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<
< |
| 1L |
1.5L |
Component |
| 1.0 ml |
1.5 ml |
10% Magnesium Sulfate MgSO4 (separately autoclaved stock) |
| 1.0 ml |
1.5 ml |
0.2% Thiamin (vitamin B1) (filter sterilized stock) |
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>
> |
| 500 mL |
1.0 L |
Component |
| 500 µL |
1.0 ml |
10% MgSO4 (separately autoclaved stock) |
| 500 µL |
1.0 ml |
0.2% Thiamine (vitamin B1) (filter sterilized stock refrigerated at 4ºC) |
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Source: Lenski Lab Protocol
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