ProtocolsPapillationAgar < Lab

---+++ PA: Lac Papillation Agar

Make 10&times; Minimal A Salts.

| *10&times; Minimal A Salts* |||
|  *1L*   |  *Component*  |  *MW*  |
|  80 g  | Potassium Phosphate (dibasic) K<sub>2</sub>HPO<sub>4</sub> |  MW 174.18 |  
|  45 g  | Potassium Phosphate (monobasic) KH<sub>2</sub>PO<sub>4</sub> | MW 136.07 |
|  10 g  | Ammonium Sulfate (NH<sub>4</sub>)<sub>2</sub> SO<sub>4</sub>| MW 132.08 |
|  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 |
|  to 1L | dH<sub>2</sub>O | |
Autoclave. Salt solution can be stored at room temperature. <br>

| *Agar Solution* ||
|  *1L*   |  *Component*  |
|  15 g  |  Agar | 
|  to 490 ml | dH<sub>2</sub>O |
Autoclave. <br>

| *Sugar Solution* ||
|  *1L*   |  *Component*  | 
|  2 g  |  Glucose | 
|  to 400 ml | dH<sub>2</sub>O |
Autoclave. <br>

After autoclaving, combine agar and sugar and add:

|  *1L*  |*Component*  |
| 100 ml | 10&times; Minimal A Salts |
| 6 ml | 10% w/v (0.083M Mg<sup>2+</sup>) Magnesium Sulfate, anhydrous <nop>MgSO<sub>4</sub> (autoclaved) MW 120.37  |
| 2.5 ml | 0.2% w/v Thiamin (vitamin B1) (filter sterilized) |
| 1 ml  | 500 mg/ml P-Gal (phenyl &beta;-D-galactoside) (filter sterilized) |
| 1 ml | 40 mg/ml X-Gal (5-bromo-4-chloro-3-indolyl &beta;-D-galactoside) (filter sterilized) |

*Note:* The amounts added of some standard components (including agar) are slightly different for this recipe. Volumes have also been altered from the original Miller recipe to use the same stock solutions we have available for making DM plates. The final concentrations are 0.2% glucose, 0.05% P-Gal, and 0.004% X-Gal (w/v).

Sources:
   1 Nghiem, Y., Cabrera, M., Cupples, C.G., Miller, J.H. (1988) The mutY gene : A mutator locus in _Escherichia coli_ that generates G•C&rarr;T•A transversions. _Proc. Natl. Acad. Sci. U.S.A._ *85*:2709-2713.
   1 Miller J.H. (1972) _Experiments in Molecular Genetics_. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
Barrick Lab > ProtocolsMediaRecipes > ProtocolsPapillationAgar
Topic revision: r2 - 2013-02-19 - 23:01:24 - Main.AurkoDasgupta