ProtocolsRecipesMinimalGlucose < Lab

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.

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

Separately, prepare the agar:

500 mL	1.0 L	Component
8 g	16 g	Agar
500 µL	1 mL	Antifoam (5%)
166.7 mL	333 mL	dH₂O

Next, prepare the sugar solution. The final concentration is 0.4% w/v:

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.

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:

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)

Source: Lenski Lab Protocol

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