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).