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 |
Copyright © 2024 Microbe Division (JCM) - All Rights Reserved