356. ANOXYNATRONUM MEDIUM

Na₂CO₃ 25.0 g

NaHCO₃ 25.0 g

KCl 0.2 g

MgCl₂ × 6 H₂O 0.1 g

NH₄Cl 0.5 g

K₂HPO₄ 0.2 g

Yeast extract 0.2 g

Trace elements SL-10 (see below) 1.0 ml

Vitamin solution (see below) 1.0 ml

Na₂S × 9 H₂O 0.7 g

Glucose 5.0 g

Distilled water 1000.0 ml, pH 9.0

Trace element solution SL-10:

HCl (25%; 7.7 M) 10.0 ml

FeCl₂ × 4 H₂O 1.5 g

ZnCl₂ 70.0 mg

MnCl₂ × 4 H₂O 100.0 mg

H₃BO₃ 6.0 mg

CoCl₂ × 6 H₂O 190.0 mg

CuCl₂ × 2 H₂O 2.0 mg

NiCl₂ × 6 H₂O 24.0 mg

Na₂MoO₄ × 2 H₂O 36.0 mg

Distilled water 990.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine-HCl 10.0 mg

Thiamine-HCl × 2 H₂O 5.0 mg

Riboflavin 5.0 mg

Nicotinic acid 5.0 mg

D-Ca-pantothenate 5.0 mg

Vitamin B₁₂ 0.1 mg

p-Aminobenzoic acid 5.0 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

 

Prepare the medium anaerobically without the vitamins, Na₂S × 9 H₂O, yeast extract, NaHCO₃, and Na₂CO₃. Boil the medium, cool under N₂, add the NaHCO₃, and Na₂CO₃, dispense under N₂ and autoclave the medium. Add the vitamin solution, yeast extract, Na₂S × 9 H₂O and glucose from sterile stock solution prepared under N₂ to the cooled, autoclaved medium.

 

Updated 1.02.2024