324. SPHAEROCHAETA MEDIUM

NaCl 1.0 g

KCl 0.5 g

MgCl26 H2O 0.4 g

CaCl22 H2O 0.1 g

NH4Cl 0.3 g

KH2PO4 0.2 g

Na2SO4 0.15 g

Xylose 20 mmol

Yeast extract 1.0 g

NaHCO3 0.5 g

Trace element solution SL-7 (see below) 1.0 ml

Vitamin solution (see below) 1.0 ml

Distilled water 990.0 ml

Trace mineral solution:

FeCl2 4 H2O 1.99 g

CoCl2 0.13 g

MnCl2 4 H2O 0.198 g

ZnCl2 0.136 g

H3BO3 0.0 62 g

NiCl2 6 H2O 0.013 g

AlCl3 6 H2O 0.0133 g

Na2MoO4 2 H2O 0.0242 g

CuCl2 2 H2O 0.0 17 g

Distilled water 1000.0 ml

Seven vitamins solution:

Vitamin B12 100.0 mg

p-Aminobenzoic acid 80.0 mg

D(+)-Biotin 20.0 mg

Nicotinic acid 200.0 mg

Calcium pantothenate 100.0 mg

Pyridoxine hydrochloride 300.0 mg

Thiamine-HCl 2 H2O 200.0 mg

Distilled water 1000.0 ml

Dissolve all components in distilled water and add 0.5 ml/l Na-resazurin solution (0.1% w/v), bring to the boil and cool to room temperature while sparging with 80% N2 and 20% CO2 gas mixture. Add L-cysteine-HCL H2O (0.3 g/l), dispense the medium into anoxic Hungate-type tubes or serum vials and autoclave. Sodium hydrogen carbonate, vitamins, xylose and antibiotics (when necessary) were added to the autoclaved medium from sterile anaerobic stock solutions. Adjust pH of medium to 7.2-7.3 with a sterile anoxic stock solution of Na2CO3 (5% w/v) prepared under 80% N2 and 20% CO2 gas atmosphere.

 

Updated 28/06/2020