Na3 citrate, 5 1/2 H20 150 grams(NOTE, you may substitute 125 g Na3 Citrate 2 H2O) KH2PO4, anhydrous 250 grams NH4NO3, anhydrous 100 grams MgSO4, 7 H20 10 grams CaCl2, 2 H20 5 grams Trace Element Solution (see below) 5 ml. Biotin Solution (see below) 2.5 ml.The resulting total volume is about 1.00 liter. Chloroform (2 ml. ) is added as a preservative.
Vogels Medium N (Minimal) is
20 mL Vogel's Salts
15 grams Sucrose
1 liter dH2O
add 15 grams agar for solid medium
The trace element solution (containing citric acid as a solubilizing agent) is made up as follows: In 95 ml. distilled water, dissolve successively with stirring at room temperature:
Citric acid, 1 H20 5. 00 grams ZnSO4, 7 H20 5. 00 grams Fe(NH4)2(SO4)2, 6 H20 1. 00 gram CuSO4, 5 H20 0. 25 gram MnSO4, 1 H20 0. 05 gram H3BO3, anhydrous 0. 05 gram Na2MoO4, 2 H20 0. 05 gramThe resulting total volume is about 100 ml. Chloroform (1 ml. ) is added as a preservative, and the trace element solution is stored at room temperature.
The biotin solution is prepared by dissolving 5.0 mg. biotin in 50 ml. distilled water. The solution obtained is dispensed in test tubes and stored in the frozen state.