141
141 .
METHANOGENIUM MEDIUM (H2/CO2)
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Medium for Methanobacterium petrolearium DSM 22353
* Modification: Adjust pH to 6.5.
Growth conditions: 3-7 days
anaerobic
35 °C
Main sol. 141 #
ml
Equipment needed: Autoclave
Filter
Hungate tubes
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KCl | 0.34 | g | 0.336 | 4.502 | |
| MgCl2 x 6 H2O | 4.00 | g | 3.949 | 19.423 | |
| MgSO4 x 7 H2O | 3.45 | g | 3.406 | 13.818 | |
| NH4Cl | 0.25 | g | 0.247 | 4.614 | |
| CaCl2 x 2 H2O | 0.14 | g | 0.138 | 0.94 | |
| K2HPO4 | 0.14 | g | 0.138 | 0.793 | |
| NaCl | 18.00 | g | 17.769 | 304.055 | |
| Modified Wolin's mineral solution | 10.00 | ml | - | - | |
|
Fe(NH4)2(SO4)2 x 6 H2O
(0.1% w/v) |
2.00 | ml | 0.002 | - | |
| Na-acetate | 1.00 | g | 0.987 | 12.034 | |
|
Yeast extract
(OXOID) |
2.00 | g | 1.974 | - | |
|
Trypticase peptone
(BD BBL) |
2.00 | g | 1.974 | - | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 4.9e-4 | 0.002 | |
| Wolin's vitamin solution | 10.00 | ml | - | - | |
| NaHCO3 | 5.00 | g | 4.936 | 58.755 | |
| L-Cysteine HCl x H2O | 0.50 | g | 0.494 | 2.81 | |
| Na2S x 9 H2O | 0.50 | g | 0.494 | 2.055 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Note: If the medium is being used without overpressure then adjust pH with a small amount of sterile anoxic 1 N HCl, if necessary. | |||||
| 2 Dissolve ingredients (except bicarbonate, vitamins, cysteine and sulfide), sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate and adjust pH to 6.5, then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. After sterilization add cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of complete medium to 6.8 - 7.0, if necessary. | |||||
| 3 For incubation use sterile 80% H2 and 20% CO2 gas mixture at two atmospheres of pressure. | |||||
Solution occurs in 166
other media (first defined in Medium 141).
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Nitrilotriacetic acid | 1.50 | g | 1.5 | 7.848 | |
| MgSO4 x 7 H2O | 3.00 | g | 3 | 12.172 | |
| MnSO4 x H2O | 0.50 | g | 0.5 | 2.958 | |
| NaCl | 1.00 | g | 1 | 17.112 | |
| FeSO4 x 7 H2O | 0.10 | g | 0.1 | 0.36 | |
| CoSO4 x 7 H2O | 0.18 | g | 0.18 | 1.161 | |
| CaCl2 x 2 H2O | 0.10 | g | 0.1 | 0.68 | |
| ZnSO4 x 7 H2O | 0.18 | g | 0.18 | 0.626 | |
| CuSO4 x 5 H2O | 0.01 | g | 0.01 | 0.04 | |
| AlK(SO4)2 x 12 H2O | 0.02 | g | 0.02 | 0.042 | |
| H3BO3 | 0.01 | g | 0.01 | 0.162 | |
| Na2MoO4 x 2 H2O | 0.01 | g | 0.01 | 0.041 | |
| NiCl2 x 6 H2O | 0.03 | g | 0.03 | 0.231 | |
| Na2SeO3 x 5 H2O | 0.30 | mg | 3.0e-4 | 0.001 | |
| Na2WO4 x 2 H2O | 0.40 | mg | 4.0e-4 | 0.001 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||
Solution occurs in 53
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 | 0.021 |
| 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 | - | - |
Metadata
| Last modified: | 21.02.24 |
| Source: | DSMZ |
| Taxonomic range: | Archaea |
| Taxonomic range: | Bacteria |
| Medium type: | Defined medium |
| Final pH: | * 6.5 |
| Equipment needed: | Autoclave, Filter, Gassing station, Hungate tubes |
| Edit in Medium builder | |
Gas composition
- 80 % H2
- 20 % CO2
Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
Molecular composition
| Compound | Conc. [g/L] | Conc. [mM] |
|---|---|---|
| NaCl | 17.769 | 304.055 |
| NaHCO3 | 4.936 | 58.755 |
| MgCl2 x 6 H2O | 3.949 | 19.423 |
| MgSO4 x 7 H2O | 3.406 | 13.818 |
| Trypticase peptone | 1.974 | - |
| Show more | ||
Cultivation metadata from 32 strains
| pH | 6 - 8 | 6 strains |
| pH | 4 - 6 | 5 strains |
| pH | 8 - 10 | 4 strains |
| phyla | 2 | 31 strains |
| oxygen | anaerobe | 30 strains |
| Show more | ||