Medium data

Search for medium no. = [556].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

556   METHYLOSOMA DIFFICILE MEDIUM

NaCl  0.1     g
MgCl2·6H2O  0.04    g
KCl  0.05    g
CaCl2·2H2O  0.015   g
Na2SO4  0.016   g
Distilled water  993.0     ml
Mix components and autoclave. After cooling, add the following solutions (autoclaved or filter-sterilized) to the medium:
2 M HEPES buffer (pH 7.0)  5.0     ml
1 M Phosphate buffer (pH 7.2, see below)  0.15    ml
0.09 M KNO3 solution  0.544   ml
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187)  1.0     ml
Vitamin solution (filter-sterilized, see below)  1.0     ml
Distribute the culture medium into culture tubes (leave >80% volume as a gas phase) and seal with butyl rubber stoppers.
Finally, replace the gas phase with 20% methane and 5% CO2 in air (by volume).
1 M Phosphate buffer:
Mix 28 ml of 1 M KH2PO4 and 78 ml of 1 M Na2HPO4 to prepare 100 ml of 1 M phosphate buffer.
Vitamin solution:
Vitamin B12  50.0     mg
p-Aminobenzoic acid  50.0     mg
Biotin  10.0     mg
Nicotinic acid  100.0     mg
DL-Calcium pantothenate  25.0     mg
Pyridoxine·HCl  250.0     mg
Thiamine·HCl  50.0     mg
Distilled water  1.0     L