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Tris-Acetate-Phosphate

Page history last edited by PBworks 14 years, 4 months ago

Tris - Acetate - Phosphate (TAP) Medium

(modified from Durnford Lab)

 

For One Liter of Medium:
2X Filner’s Beijernicks Solution25ml
1M Potassium Phosphate1ml
Trace mineral solution or5ml
P IV solution 2X (see SVM)3ml
Tris-Base2.42g
Glacial Acetic Acid (17.4 mM acetate)1ml
(pH should be 7.2)

 

Stock solutions:

 

2X Filner’s Beijernicks Solution (500 ml)

NH4Cl8g
CaCl2 * 2H2O1g
MgSO4 *7H2O2g

 

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 gFeSO4*7H2O
2.2 gZnSO4*7H2O
1.14 gH3BO3
0.51 gMnCl2*4H2O
0.016 gCuSO4*5H2O
0.073 gNa2MoO4*2H2O
0.016 gCoCl2*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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