(r3) ProtocolsRecipesDavisMingioli < Lab

---+++ DM: Davis Minimal

*Notes:  Salts, agar and sugars need to be prepared and autoclaved separately.* <br>

For making 1.5 liters of DM agar do the following: <br>

Combine the following in a 2 L flask (including magnetic stir bar):  <br>

| 10.5 g | Potassium Phosphate (dibasic, trihydrate) HK<sub>2</sub>PO<sub>4</sub> (H<sub>2</sub>O)<sub>3</sub> | MW 228.17 | 
| 3 g | Potassium Phosphate (monobasic) H<sub>2</sub>KPO<sub>4</sub> | MW 136.07 |
| 1.5 g | Ammonium Sulfate (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>| MW 132.08 |
| 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 |
Add 500 ml of dH<sub>2</sub>O, cover with tin foil and autoclave. <br>

Next, prepare the agar in a 1 liter flask (including magnetic stir bar) by adding the following: <br>

| 24 g | Agar |
| 1.5 ml | Antifoam (5%) |

Add 500 ml of dH<sub>2</sub>O, cover with tin foil and autoclave. <br>

Next, prepare the sugar in a separate 1 liter flask by adding the following: <br>

| 6 g | Glucose (or other sugar) |

Add 500 ml of dH<sub>2</sub>O, cover with tin foil and autoclave. <br>

After autoclaving combine the contents of the three flasks into the larger flask and add the following stock solutions: <br>


| 1.5 ml | 10% Magnesium Sulfate MgSO<sub>4</sub> | MW 120.37 | | 
| 1.5 ml | 0.2% Thiamin (vitamin B1) | Note* Thiamin must be syringe filtered into the media | |


To make 1 liter of DM plates combine the following in a 2 L flask (including magnetic stir bar):  <br>

| 7 g | Potassium Phosphate (dibasic, trihydrate) HK<sub>2</sub>PO<sub>4</sub> (H<sub>2</sub>O)<sub>3</sub> | MW 228.17 | 
| 2 g | Potassium Phosphate (monobasic) H<sub>2</sub>KPO<sub>4</sub> | MW 136.07 |
| 1 g | Ammonium Sulfate (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>| MW 132.08 |
| 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>| MW 294.10 |
Add about 333 ml of dH<sub>2</sub>O, cover with tin foil and autoclave. <br>

Next, prepare the agar in a 1 liter flask (including magnetic stir bar) by adding the following: <br>

|16 g | Agar |
| 1 ml | Antifoam (5%) |

Add about 333 ml of dH<sub>2</sub>O, cover with tin foil and autoclave. <br>

Next, prepare the sugar in a separate 1 liter flask by adding the following: <br>

| 4 g | Glucose (or other sugar) |

Add about 333 ml of dH<sub>2</sub>O, cover with tin foil and autoclave. <br>

After autoclaving combine the contents of the three flasks into the larger flask and add the following stock solutions: <br>


| 1 ml | 10% Magnesium Sulfate MgSO<sub>4</sub> | MW 120.37 | | 
| 1 ml | 0.2% Thiamin (vitamin B1) |Note* Thiamin must be syringe filtered into the media | |


Final nutrient concentrations:
| 67.4 mM | Phosphate (PO<sub>4</sub><sup>3-</sup>) |
| 8.38 mM | Sulfate (SO<sub>4</sub><sup>2-</sup>) |
| 15.1 mM | Ammonium (NH<sub>4</sub>) | 
| 5.1 mM | Sodium (Na<sup>+</sup>) |
| 98.1 mM | Potassium (K<sup>+</sup>) |
| 0.83 mM | Magnesium (MGgsup>2+</sup>) |
| 1.7 mM | Citrate |
| 139 µM | Glucose (in DM25) |
Barrick Lab > ProtocolsMediaRecipes > ProtocolsRecipesDavisMingioli
Topic revision: r3 - 2011-07-26 - 12:50:08 - Main.CraigBarnhart