Medium data

Search for medium no. = [251].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

251   METHANOTHERMUS FERVIDUS MEDIUM

Mineral solution I (see below)  37.5     ml
Mineral solution II (see below)  37.5     ml
(NH4)2Ni(SO4)2·6H2O  3.0     mg
FeSO4·7H2O  2.0     mg
Yeast extract (BD-Difco)  2.0     g
Trypticase peptone (BD-BBL)  2.0     g
Trace vitamins (see Medium No. 197)  10.0     ml
Trace minerals (see Medium No. 151)  10.0     ml
Na2SO4  3.4     g
NaHCO3  2.0     g
Resazurin  1.0     mg
Na2S·9H2O  0.5     g
L-Cysteine·HCl·H2O  0.5     g
Distilled water  920.0     ml
Mix ingredients, except NaHCO3, L-cysteine·HCl·H2O and Na2S·9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of H2-CO2 (80:20), and add NaHCO3 to the medium. Dispense the medium into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight and adjust pH to 6.5, if necessary. Separately autoclave L-cysteine·HCl and Na2S·9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the L-cysteine·HCl and Na2S·9H2O solutions. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20).
Mineral solution I:
K2HPO4  6.0     g
Distilled water  1.0     L
Mineral solution II:
KH2PO4  6.0     g
(NH4)2SO4  6.0     g
NaCl  12.0     g
MgSO4·7H2O  2.4     g
CaCl2·2H2O  1.6     g
Distilled water  1.0     L