Barrick Lab :: ProtocolsRecipesMinimalGlucose

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---+++ MG:  Minimal Glucose agar, aka DM: Davis Mingioli agar
*%RED%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 [[http://schaechter.asmblog.org/schaechter/2014/12/the-great-plate-count-anomaly-that-is-no-more.html][agar and phosphate]] or phosphate and glucose are autoclaved together.%ENDCOLOR%*

|  *500 mL*  |  *1.0L*  | *Component* |
|  2.65 g  |  5.3 g | Potassium Phosphate (dibasic) K<sub>2</sub>HPO<sub>4</sub> | 
|  1 g  |  2 g  | Potassium Phosphate (monobasic) KH<sub>2</sub>PO<sub>4</sub> |
|  0.5 g  |  1 g  | Ammonium Sulfate (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>|
|  0.25 g  |  0.5 g  | Sodium Citrate (trisodium, dihydrate) Na<sub>3</sub>C<sub>6</sub>H<sub>5</sub>O<sub>7</sub>  (H<sub>2</sub>O)<sub>2</sub>|
|  166.7 mL  |  333 mL  | dH<sub>2</sub>O |

Separately, prepare the agar: <br>

|  *500 mL*  |  *1.0 L*  | *Component* | 
|  8 g  |  16 g  | Agar |
|  500 µL  |  1 mL  | Antifoam (5%) |
|  166.7 mL  |  333 mL  | dH<sub>2</sub>O |

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

|  *500 mL*  |  *1.0 L*  | *Component* | 
|  2 g  |  4 g  | Glucose, arabinose, or other sugar |
|  166.7 mL  |  333 mL  | dH<sub>2</sub>O |
*Autoclave the 3 parts separately for a 40 minute sterilization time.* <br>

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: <br>

|  *500 mL*  |  *1.0 L*  | *Component* |
|  500 µL  |  1.0 ml  | 10% MgSO<sub>4</sub> (separately autoclaved stock)  |
|  500 µL  |  1.0 ml  | 0.2% Thiamine (vitamin B1) (filter sterilized stock refrigerated at 4ºC) |

Source: [[http://myxo.css.msu.edu/ecoli/dmagar.html][Lenski Lab Protocol]]

Barrick Lab > ProtocolsMediaRecipes > ProtocolsRecipesMinimalGlucose

Contributors to this topic

JeffreyBarrick, CraigBarnhart, AlexaMorton, AurkoDasgupta

Topic revision: r12 - 2025-01-09 - 20:14:37 - Main.AlexaMorton