Medium data

Search for medium no. = [600].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

600   G5G6 MEDIUM

Solution A:
Na2HPO4·12H2O  0.5     g
KH2PO4  0.65    g
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see below)  1.0     ml
Resazurin  0.5     mg
Distilled water  880.0     ml
Solution B:
CaCl2·2H2O  0.11    g
MgCl2·6H2O  0.1     g
Distilled water  50.0     ml
Solution C:
NH4HCO3  0.44    g
NaHCO3  3.73    g
L-Cysteine·HCl·H2O  0.175   g
Distilled water  50.0     ml
Prepare all solutions and autoclave under a N2 atmosphere. After cooling, combine the solutions and replace the gas phase with a N2-CO2 (4:1, v/v) gas mixture. Then, add the following autoclaved or *filter-sterilized solutions to the medium:
1.0 M NaNO3 solution  10.0     ml
Trace vitamins* (see Medium No. 197)  10.0     ml
Vitamin B12 solution* (see Medium No. 403)  1.0     ml
Check the final pH to be around 7.3.
Distribute appropriate amount of the medium into culture vessels under the same gas mixture (e.g., 40.0 ml in 120 ml serum bottle) and seal with Viton rubber stoppers. Finally add per liter the following solution from an anaerobic stock (autoclaved):
5.0 mM Toluene (water-saturated)  40.0     ml
Trace element solution:
Trace element solution (see Medium No. 187)  1.0     L
AlCl3·6H2O  10.0     mg
Comment: For strain JCM 14632, incubate with a 5-10% inoculum. When toluene is depleted, re-add the toluene solution (normally every 3 to 4 days, until enough cell growth is seen).



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