Culture medium recipe
Vol.
×
Main sol. 1020 #
ml
Equipment needed: Autoclave
Hungate tubes
Filter
Gassing station
Serum bottle
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| NaNO3 | 0.2 | g | 0.2 | 2.348 | |
| KH2PO4 | 0.2 | g | 0.2 | 1.467 | |
| NH4Cl | 0.2 | g | 0.2 | 3.732 | |
| MgCl2 x 6 H2O | 0.4 | g | 0.399 | 1.964 | |
| KCl | 0.2 | g | 0.2 | 2.677 | |
| CaCl2 x 2 H2O | 0.1 | g | 0.1 | 0.679 | |
| Trace element solution SL-4 | 2.0 | ml | - | - | |
| Na2S2O3 x 5 H2O | 2.5 | g | 2.495 | 10.053 | |
| Distilled water | 1000.0 | ml | - | - | |
| 1 Dissolve ingredients, except sodium thiosulfate, then sparge solution with 80% N2 and 20% CO2 gas mixture for 30 - 45 minto make it anoxic. Adjust pH to 7.0, then distribute medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml bottles) and autoclave. Prior to inoculation, add thiosulfate from an anoxic stock solution prepared under 100% N2 gas and sterilized by filtration. Adjust pH of complete medium to 7.0, if necessary. After inoculation, pressurize culture vessels to 0.5 bar overpressure with sterile 100% H2 gas. | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Na2-EDTA | 0.50 | g | 0.5 | 1.487 | |
| FeSO4 x 7 H2O | 0.20 | g | 0.2 | 0.719 | |
| ZnSO4 x 7 H2O | 0.10 | g | 0.1 | 0.348 | |
| MnCl2 x 4 H2O | 0.03 | g | 0.03 | 0.152 | |
| H3BO3 | 0.30 | g | 0.3 | 4.851 | |
| CoCl2 x 6 H2O | 0.20 | g | 0.2 | 1.54 | |
| CuCl2 x 2 H2O | 0.01 | g | 0.01 | 0.059 | |
| NiCl2 x 6 H2O | 0.02 | g | 0.02 | 0.154 | |
| Na2MoO4 x 2 H2O | 0.03 | g | 0.03 | 0.124 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. | |||||
No comments for this medium.