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Main sol. J1221

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Internal Solution-ID: S5349

Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
MgSO4 x 7 H2O 1.00 g 0.984 3.993
CaCl2 x 2 H2O 0.20 g 0.197 1.339
KNO3 0.25 g 0.246 2.434
FeCl2 solution 1.00 ml - -
Trace element solution 1.00 ml - -
HEPES
(pH 7.0, 1 M) 		2.00 ml - -
Ammonium ferric citrate 4.00 mg 0.004 0.015
Distilled water 1000.00 ml - -
1 Mix components thoroughly and autoclave. After cooling, add the following solutions (autoclaved) to the medium:
Phosphate buffer solution 2.00 ml - -
Vitamin solution 10.00 ml - -
2 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.
3 Adjust pH to 6.8, if necessary. Cultivate with methane or methanol as a substrate as described below:
4 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.
J1221 MODIFIED NITRATE MINERAL SALTS MEDIUM-2