Barrick Lab :: ProtocolsRecipesMinimalGlucose

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	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

Separately, prepare the agar:

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

Autoclave.

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

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.

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:

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)

Source: Lenski Lab Protocol

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Contributors to this topic

JeffreyBarrick, CraigBarnhart, AlexaMorton, AurkoDasgupta

Topic revision: r11 - 2024-10-22 - 14:00:59 - Main.AlexaMorton