Culture medium recipe
Vol.
×
Main sol. 1508 #
ml
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
Sea Salt
(SIGMA) |
21.00 | g | 21 | - | |
| K2HPO4 | 0.10 | g | 0.1 | 0.574 | |
| NH4Cl | 0.10 | g | 0.1 | 1.87 | |
| NaNO3 | 0.05 | g | 0.05 | 0.588 | |
| Yeast extract | 0.05 | g | 0.05 | - | |
| Fe(III) citrate | 0.01 | g | 0.01 | 0.041 | |
| Trace element solution SL-4 | 1.00 | ml | - | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
| Na2CO3 | 1.50 | g | 1.5 | 14.153 | |
| Na-pyruvate | 1.00 | g | 1 | 9.087 | |
| Wolin's vitamin solution | 10.00 | ml | - | - | |
| Na2S2O3 x 5 H2O | 1.00 | g | 1 | 4.029 | |
| Na2S x 9 H2O | 0.25 | g | 0.25 | 1.041 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients except carbonate, pyruvate, vitamins, thiosulfate and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 minto make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add pyruvate, vitamins, thiosulfate and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of pyruvate, vitamins and thiosulfate should be sterilized by filtration. Prior to use adjust pH of complete medium to 7.3 - 7.5, if necessary. | |||||
| 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. | |||||
Solution occurs in 351
other media (first defined in Medium 141).
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| Biotin | 2.0 | mg | 0.002 | 0.008 |
| Folic acid | 2.0 | mg | 0.002 | 0.005 |
| Pyridoxine hydrochloride | 10.0 | mg | 0.01 | 0.049 |
| Thiamine HCl | 5.0 | mg | 0.005 | 0.015 |
| Riboflavin | 5.0 | mg | 0.005 | 0.013 |
| Nicotinic acid | 5.0 | mg | 0.005 | 0.041 |
| Calcium D-(+)-pantothenate | 5.0 | mg | 0.005 | - |
| Vitamin B12 | 0.1 | mg | 1.0e-4 | 7.4e-5 |
| p-Aminobenzoic acid | 5.0 | mg | 0.005 | 0.036 |
| (DL)-alpha-Lipoic acid | 5.0 | mg | 0.005 | 0.024 |
| Distilled water | 1000.0 | ml | - | - |
No comments for this medium.