Medium data

Search for medium no. = [1221].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.

1221   MODIFIED NITRATE MINERAL SALTS MEDIUM-2

MgSO4·7H2O  1.0     g
CaCl2·2H2O  0.2     g
KNO3  0.25    g
FeCl2 solution (see Medium No. 187)  1.0     ml
Trace element solution (see Medium No. 187)  1.0     ml
1 M HEPES solution (pH 7.0)  2.0     ml
Ammonium ferric citrate  4.0     mg
Distilled water  1.0     L
Mix components thoroughly and autoclave. After cooling, add the following solutions (autoclaved) to the medium:
Phosphate buffer solution (see below)  2.0     ml
Vitamin solution (see below)  10.0     ml
Adjust pH to 6.8, if necessary. Cultivate with methane or methanol as a substrate as described below:
Methane: Aseptically distribute the medium into sterile serum bottles (leave ≥ 80% gas phase by volume) and seal with rubber stoppers. Replace the gas phase with an air-methane gas mixture (75:25, v/v). Cultivate with shaking.
Methanol: Aseptically distribute the medium into culture vessels (e.g., 20 ml in 60 ml serum bottles), supplement with 4.0 ml per liter filter-sterilized methanol and seal with rubber stoppers.
Phosphate buffer solution:
KH2PO4  1.4     g
Na2HPO4·2H2O  3.6     g
Distilled water  100.0     ml
Vitamin solution:
Trace vitamins (see Medium No. 197)  1.0     L
Vitamin B12  20.0     mg
L-Ascorbic acid  5.0     mg
Nicotinamide  5.0     mg