Tris - Acetate - Phosphate (TAP) Medium
(modified from Durnford Lab)
For One Liter of Medium: |
2X Filner’s Beijernicks Solution | 25ml |
1M Potassium Phosphate | 1ml |
Trace mineral solution or | 5ml |
P IV solution 2X (see SVM) | 3ml |
Tris-Base | 2.42g |
Glacial Acetic Acid (17.4 mM acetate) | 1ml |
(pH should be 7.2) |
Stock solutions:
2X Filner’s Beijernicks Solution (500 ml)
NH4Cl | 8g |
CaCl2 * 2H2O | 1g |
MgSO4 *7H2O | 2g |
Add deionized/distilled water to 500ml and Autoclave: Store at 4°C
Trace Mineral Solution (500 ml)
5 g disodium EDTA – dissolve in 400 ml water by heating and stirring
Neutralize to pH 6.5 with 5N NaoH
Add each of the following in order. Allow each to dissolve completely before adding the next.
0.5 g | FeSO4*7H2O |
2.2 g | ZnSO4*7H2O |
1.14 g | H3BO3 |
0.51 g | MnCl2*4H2O |
0.016 g | CuSO4*5H2O |
0.073 g | Na2MoO4*2H2O |
0.016 g | CoCl2*6H2O |
Bring volume up to 500 ml and autoclave. The solution should be pale green, turning purple upon storage.
1M Potassium Phosphate Stock (50 ml)
add:
20 ml 1M stock KH2PO4 | (1M stock: 6.8g/50ml) |
30 ml 1M stock K2HPO4 | (1M stock: 8.7g/50ml) |
pH 7.2 at RT
For TAP plates or slants, add 1.3g AGAR per 100 ml TAP medium.
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