169. HETEROTROPHIC MEDIUM H3P

Solution 1:

KH₂PO₄ 2.3 g

Na₂HPO₄ × 2 H₂O 2.9 g

Distilled water 50.0 ml

Solution 2:

NH₄Cl 1.0 g

MgSO₄ × 7 H₂O 0.5 g

CaCl₂ × 2 H₂O 0.01 g

MnCl₂ × 4 H₂O 0.0 5 g

NaVO₃ × H₂O 0.0 5 g

Trace element solution SL-6 (see below) 5.0 ml

Agar (if necessary) 15.0 g

Distilled water 850.0 ml

Solution 3:

Fe(NH₄)-citrate 0.05 g

Distilled water 20.0 ml

Solution 4:

Yeast extract 1.0 g

Na-acetate × 3 H₂O 1.0 g

Na₂-succinate 1.0 g

DL-Malate 1.0 g

Distilled water 30.0 ml

pH 7.0

Solution 5:

Na-lactate 1.0 g

Na-pyruvate 1.0 g

D-Mannitol 1.0 g

D-Glucose 2.0 g

Distilled water 50.0 ml

pH 7.0

Trace element solution SL-6:

ZnSO₄ × 7 H₂O 0.1 g

MnCl₂ × 4 H₂O 0.03 g

H₃BO₃ 0.3 g

CoCl₂ × 6 H₂O 0.2 g

CuCl₂ × 2 H₂O 0.01 g

NiCl₂ × 6 H₂O 0.02 g

Na₂MoO₄ 0.03 g

Distilled water 1000.0 ml

vitamin solution:

Riboflavin 10.0 mg

Thiamine-HCl × 2 H₂O 50.0 mg

Nicotinic acid 50.0 mg

Pyridoxine-HCl 50.0 mg

Ca-pantothenate 50.0 mg

Biotin 0.1 mg

Folic acid 0.2 mg

Vitamin B₁₂ 1.0 mg

Distilled water 100.0 ml

Solutions 1, 2, 3, 4 and trace element solution are autoclaved separately for 15 min at 121°C, cooled down to 50°C and then mixed aseptically with filter-sterilized solution 5 (at 50°C) and 5.0 ml of filter-sterilized standard vitamin solution. The final pH of this medium should be 6.8 without adjustment.

 

Updated 1.02.2024