360. METHANOSARCINA MEDIUM

NaCl 5.0 g

MgCl2 6 H2O 0.2 g

CaCl2 2 H2O 0.1 g

NH4Cl 1.0 g

Casamino acids 1.0 g

Resazurin 0.01 g

Methanol 10% 10.0 ml

Trace element solution (see below) 10.0 ml

Vitamin solution (see below) 5.0 ml

L-Cysteine-HCl H2O 0.5 g

Distilled water 965.0 ml

Buffer solutions:

a) K2HPO4 29.0 g

Distilled water 100.0 ml

b) KH2PO4 15.0 g

Distilled water 100.0 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

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

pH 7.2-7.4

Prepare medium in anaerobic conditions, blowing through with N2 without O2 up to sterilization. Solutions of buffer (1 ml) add to base medium after separate sterilization. Base medium, trace element and buffer solutions autoclave at 121C for 15 min. Vitamin solution is filter sterilized.

 

Updated 11.07.2021