Medium data

Search for medium no. = [682].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

682   METHANOFOLLIS ETHANOLICUS MEDIUM

Solution A:
Yeast extract (BD-Difco)  0.1     g
KH2PO4  0.14    g
MgCl2·6H2O  0.2     g
CaCl2·2H2O  0.15    g
NH4Cl  0.54    g
Trace vitamins solution (see below)  2.0     ml
Trace element solution (see below)  1.0     ml
NaHCO3  2.5     g
Resazurin  1.0     mg
Distilled water  900.0     ml
Mix components, except NaHCO3, and adjust pH to 6.5. Bring to a boil for 5-10 sec., then, cool down under a stream of N2-CO2 (80:20, v/v) and add NaHCO3 to the solution. Dispense 0.9 volume of the medium into culture vessels (e.g., 18 ml of the medium in 50 ml serum bottle) under a stream of N2-CO2 (80:20, v/v) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight.
Solution B:
Ethanol  0.46    ml
Distilled water  100.0     ml
Adjust pH to 7.0 and filter-sterilize. Keep the solution under a N2 atmosphere.
To complete the medium, add 0.1 volume of solution B to the culture vessels containing solution A. Prior to inoculation, aseptically and anaerobically add 0.01 volume of L-cysteine·HCl·H2O and Na2S·9H2O solutions to the medium (3% solutions, autoclaved and stored under a N2 atmosphere). Incubate with a 10% inoculum.
Trace vitamins solution:
Biotin  4.9     mg
Folic acid  8.8     mg
Pyridoxine·HCl  4.1     mg
Thiamine·HCl  6.7     mg
Riboflavin  7.5     mg
Nicotinic acid  2.5     mg
DL-Calcium pantothenate  9.5     mg
Vitamin B12  0.1     mg
p-Aminobenzoic acid  2.7     mg
Lipoic acid  4.1     mg
Distilled water  1.0     L
Trace element solution:
FeCl3·6H2O  1.27    g
MnCl2·4H2O  0.2     g
CoCl2·6H2O  0.13    g
ZnCl2  0.14    g
CuCl2·2H2O  0.001   g
AlCl3  0.01    g
H3BO3  0.006   g
Na2MoO4·2H2O  0.02    g
NiCl2·6H2O  0.01    g
Na2SeO3  0.002   g
Na2WO4·2H2O  0.003   g
Distilled water  1.0     L