Medium data
Search for medium no. = [403].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
403 ACETOBACTERIUM SUBMARINUS MEDIUM
| Sea salts (Sigma) | 30.0 | g |
| Yeast extract (BD-Difco) | 0.1 | g |
| Trace element solution (see Medium No. 301) | 1.0 | ml |
| Vitamin solution (see below) | 1.0 | ml |
| Thiamine solution (see below) | 1.0 | ml |
| Vitamin B12 solution (see below) | 1.0 | ml |
| Na2WO4·2H2O | 30.0 | mg |
| Na2SeO4 | 0.5 | mg |
| Sodium acetate | 1.0 | g |
| Methylamine | 2.0 | g |
| Sodium formate | 5.0 | g |
| Methanol | 5.0 | ml |
| KH2PO4 | 0.2 | g |
| NH4Cl | 0.27 | g |
| Resazurin | 0.5 | mg |
| NaHCO3 | 1.26 | g |
| Na2S·9H2O | 0.5 | g |
| Distilled water | 1.0 | L |
Mix ingredients except methanol, NaHCO3 and Na2S·9H2O, bring to a boil and cool down under a stream of N2 gas. Add methanol to the medium and distribute into culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers, and autoclave. Separately autoclave 5% Na2S·9H2O solution under a N2 atmosphere. Filer-sterilize 8% NaHCO3 solution and store under a N2-CO2 (4:1, v/v) gas mixture. Before inoculation, add the sterile NaHCO3 and Na2S·9H2O solutions to the medium, and readjust pH to 7.0, if necessary. Pressurize the inoculated bottles to 200 kPa H2-CO2 (4:1, v/v).
Vitamin solution:
| p-Aminobenzoic acid | 4.0 | mg |
| Biotin | 1.0 | mg |
| Nicotinic acid | 10.0 | mg |
| DL-Calcium pantothenate | 5.0 | mg |
| Pyridoxine·HCl | 10.0 | mg |
| Sodium phosphate buffer (10 mM, pH 7.1) | 100.0 | ml |
Thiamine solution:
| Thiamine·HCl | 10.0 | mg |
| Sodium phosphate buffer (25 mM, pH 3.4) | 100.0 | ml |
Vitamin B12 solution:
| Vitamin B12 | 5.0 | mg |
| Distilled water | 100.0 | ml |
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