241. OCEANITHERMUS PROFUNDUS MEDIUM

NH4Cl 0.33 g

MgCl2 6 H2O 0.33 g

CaCl2 2 H2O 0.33 g

KCl 0.33 g

KNO3 0.33 g

NaCl 30.0 g

HEPES 2.38 g

Yeast extract 0.2 g

Tryptone 1.0 g

Sucrose 2.0 g

Vitamin solution (see below) 1.0 ml

Trace elements (see below) 1.0 ml

Distilled water 1000.0 ml

Vitamin solution:

Biotin 2.0 mg

Folic acid 2.0 mg

Pyridoxine 10.0 mg

Riboflavin 5.0 mg

Pantotenoic acid 5.0 mg

p-Aminobenzoic acid 5.0 mg

Thiamine-HCl 5.0 mg

Nicotinic acid 5.0 mg

Vitamin B12 0.1 mg

Lipoic acid 5.0 mg

Distilled water 1000.0 ml

Trace element solution:

Nitrilotriacetic acid 12.8 g

FeSO4 7 H2O 0.1 g

MnCl2 4 H2O 0.1 g

CoCl2 2 H2O 0.17 g

CaCl2 2 H2O 0.1 g

ZnCl2 0.1 g

CuCl2 2 H2O 0.02 g

H3BO3 0.01 g

Na2MoO4 2 H2O 0.01 g

NaCl 1.0 g

Na2SeO4 0.017 g

Distilled water 1000.0 ml

Prepare the medium anaerobically, under N2, omitting the CaCl2 2 H2O, MgCl2 6 H2O, KNO3, tryptone, yeast extract, vitamins and sucrose. The pH should be 7.0-7.5. Dispense the medium into vessels suitable for anaerobic growth (Hungate tubes or serum bottles) under an atmosphere of N2 and autoclave at 121C 15 min. To the sterile, cooled medium add, from sterile stock solutions the CaCl2 2 H2O, MgCl2 6 H2O, KNO3, tryptone, yeast extract, vitamins and sucrose. The CaCl2 2 H2O, MgCl2 6 H2O, KNO3, tryptone, and yeast extract, stock solutions should be autoclaved at 121C 15 min, sucrose at 111 30 min, while the vitamin solution is sterile filtered.

 

Updated 24.06.2021