Medium data
Search for medium no. = [944].
Unless otherwise stated, sterilize media by autoclaving at 121°C for 15 min.
944 MW MEDIUM FOR METHANOSARCINA
| NaCl | 1.0 | g |
| KCl | 0.5 | g |
| MgCl2·6H2O | 0.4 | g |
| NH4Cl | 0.25 | g |
| CaCl2·2H2O | 0.15 | g |
| Trace elements solution (see below) | 10.0 | ml |
| Resazurin | 1.0 | mg |
| Distilled water | 946.0 | ml |
Mix components thoroughly, and autoclave under a N2-CO2 (80:20, v/v) gas atmosphere. After cooling, aseptically and anaerobically add the following solutions (autoclaved or *filter-sterilized):
| 10% KH2PO4 solution | 2.0 | ml |
| 8% NaHCO3 solution* | 32.0 | ml |
| Vitamin solution* (see below) | 10.0 | ml |
Aseptically distribute the medium into culture vessels under a H2-CO2 (80:20, v/v) gas mixture and seal with butyl rubber stoppers (leave >80% gas phase by volume). Finally, add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N2 atmosphere):
| 5% L-Cysteine·HCl·H2O solution | 6.0 | ml |
| 5% Na2S·9H2O solution | 6.0 | ml |
Pressurize the inoculated vessels to 200 kPa H2-CO2 (80:20, v/v).
Trace elements solution:
| Nitrilotriacetic acid | 1.5 | g |
| Fe(NH4)2(SO4)2·6H2O | 0.2 | g |
| Na2SeO3·5H2O | 0.2 | g |
| MnCl2·4H2O | 0.11 | g |
| CoCl2·6H2O | 0.1 | g |
| Na2MoO4·2H2O | 0.1 | g |
| NaWO4·2H2O | 0.1 | g |
| NiCl2·6H2O | 0.25 | g |
| ZnSO4·7H2O | 0.2 | g |
| H3BO3 | 0.01 | g |
| CuSO4·5H2O | 0.002 | g |
| Distilled water | 1.0 | L |
Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0.
Vitamin solution:
| p-Aminobenzoic acid | 10.0 | mg |
| Nicotinic acid | 10.0 | mg |
| DL-Calcium pantothenate | 10.0 | mg |
| Pyridoxine·HCl | 10.0 | mg |
| Riboflavin | 10.0 | mg |
| Thiamine·HCl | 10.0 | mg |
| Biotin | 5.0 | mg |
| Folic acid | 5.0 | mg |
| Lipoic acid | 5.0 | mg |
| Vitamin B12 | 5.0 | mg |
| Distilled water | 1.0 | L |
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