ProtocolsRecipesMinimalGlucose < Lab

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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*  |  *1L*  |  *1.5L*  |  *Component*  |  *MW*  |
|  2.65 g  |  5.3 g  |  8 g  | Potassium Phosphate (dibasic) K<sub>2</sub>HPO<sub>4</sub> | MW 174.18 |  
|  1 g  |  2 g  |  3 g  | Potassium Phosphate (monobasic) KH<sub>2</sub>PO<sub>4</sub> | MW 136.07 |
|  0.5 g  |  1 g  |  1.5 g  | Ammonium Sulfate (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>| MW 132.08 |
|  0.25 g  |  0.5 g  |  0.75 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>| MW 294.10 |
|  166.7 ml  |  333 ml  |  500 ml  | dH<sub>2</sub>O | |

Separately, prepare the agar: <br>

|  *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 | dH<sub>2</sub>O |
*Autoclave.* <br>

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

|  *500 ml*  |  *1L*  |  *1.5L*  |  *Component*  | 
|  2 g  |  4 g  |  6 g  | Glucose, arabinose, or other sugar |
|  166.7 ml  |  333 ml  |  500 ml  | dH<sub>2</sub>O |
*Autoclave.* <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>

|  *1L*  |  *1.5L*  |  *Component*  |
| 1.0 ml | 1.5 ml | 10% Magnesium Sulfate MgSO<sub>4</sub> (separately autoclaved stock)  |
| 1.0 ml | 1.5 ml | 0.2% Thiamin (vitamin B1) (filter sterilized stock) |

Source: [[http://myxo.css.msu.edu/ecoli/dmagar.html][Lenski Lab Protocol]]
This topic: Lab > ProtocolsMediaRecipes > ProtocolsRecipesMinimalGlucose
Topic revision: r11 - 2024-10-22 - AlexaMorton