Medium data

Search for medium no. = [1246].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1246   BICARBONATE-BUFFERED MEDIUM WITH ACETHYLENE

CaCl2·2H2O  0.225   g
KCl  0.2     g
MgCl2·6H2O  0.2     g
KH2PO4  0.02    g
1% FeSO4·6H2O solution (in 0.02 N H2SO4)  0.1     ml
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187)  1.0     ml
NaCl  20.0     g
NH4Cl  0.2     g
HEPES  5.2     g
Distilled water  1.0     L
Mix components thoroughly and adjust pH to 7.5. Distribute the medium in culture vessels (e.g., 10 ml in Balch-type tubes), flush with N2 and seal with butyl rubber stoppers. Autoclave. After cooling, add per liter the following solution (autoclaved or *filter-sterilized):
8% NaHCO3 solution*  20.0     ml
5% L-Cysteine·HCl·H2O solution  5.0     ml
After inoculation, add sterile pure acetylene gas, made from calcium carbide and water, to the gas phase (the first time, 0.5% volume of the gas phase: the next day, 1.0% volume: increase each day until 2.5% volume is reached).