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

0.16 g           CuSO₄*5H₂O

0.073 g           Na₂MoO₄*2H₂O

0.16 g           CoCl₂*6H₂O

 

Bring volume up to 500 ml and autoclave. The solution should be pale gree, 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.