| Na2SO4 | 3 | g |
| NH4Cl | 0.3 | g |
| KH2PO4 | 0.2 | g |
| MgCl2·6H2O | 3.1 | g |
| CaCl2·2H2O | 0.15 | g |
| NaCl | 21 | g |
| KCl | 0.5 | g |
| Trace element solution* | 1 | ml |
| Selenite-tungstate solution** | 1 | ml |
| Resazurin | 1 | mg |
| Na2CO3 | 2.5 | g |
| Sodium lactate | 2.5 | g |
| Vitamin solution*** | 10 | ml |
| Na2S·9H2O | 0.4 | g |
| Distilled water | 990 | ml |
|
| | Mix ingredients (except Na2CO3, sodium lactate, vitamin solution and Na2S·9H2O) and dispense into suitable culture vessels under a stream of N2/CO2 (80/20) gas, seal with butyl rubber stoppers, and then autoclave. Separately autoclave Na2CO3 under a N2/CO2 (80/20) atmosphere. Separately autoclave sodium lactate and Na2S·9H2O under a N2 atmosphere. Sterilize vitamin solution by filtration. To complete the medium, add Na2CO3, sodium lactate, vitamin solution and Na2S·9H2O. Adjust pH of the complete medium to around 7.5, if necessary. |
|
| | Addition of 10-20 mg/L sodium dithionite may stimulate growth at the begining. |
| | | | | | |
| | *Trace element solution |
| | | HCl (35%) | 7 | ml |
| | | FeCl3·6H2O | 1.5 | g |
| | | ZnCl2 | 70 | mg |
| | | MnCl2·4H2O | 100 | mg |
| | | H3BO3 | 6 | mg |
| | | CoCl2·6H2O | 190 | mg |
| | | CuCl2·2H2O | 2 | mg |
| | | NiCl2·6H2O | 24 | mg |
| | | Na2MoO4·2H2O | 36 | mg |
| | | Distilled water | 1 | L |
|
| | First dissolve FeCl3 in conc. HCl, and dilute in water, then add the other minerals. |
| | | | | | |
| | **Selenite-tungstate solution |
| | | NaOH | 0.5 | g |
| | | Na2SeO3·5H2O | 3 | mg |
| | | Na2WO4·2H2O | 4 | mg |
| | | Distilled water | 1 | L |
| | | | | | |
| | ***Vitamin solution |
| | | Biotin | 2 | mg |
| | | Folic acid | 2 | mg |
| | | Pyridoxine-HCl (VitaminB6) | 10 | mg |
| | | Thiamine-HCl·H2O (Vitamin B1) | 5 | mg |
| | | Riboflavin (Vitamine B2) | 5 | mg |
| | | Nicotinic acid | 5 | mg |
| | | D-Ca-pantothenate | 5 | mg |
| | | Vitamin B12 | 0.1 | mg |
| | | p-Aminobenzoic acid | 5 | mg |
| | | Lipoic acid | 5 | mg |
| | | Distilled water | 1 | L |
|
| | Adjust pH to 7.0 with NaOH. |