1523
1523 .
MODIFIED METHANOBACTERIUM MEDIUM
Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Culture medium recipe
Vol.
×
Main sol. 1523 #
ml
Equipment needed: Hungate tubes
Filter
Autoclave
Gassing station
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| KH2PO4 | 0.50 | g | 0.499 | 3.667 | |
| MgSO4 x 7 H2O | 0.40 | g | 0.399 | 1.62 | |
| NaCl | 0.40 | g | 0.399 | 6.831 | |
| NH4Cl | 0.40 | g | 0.399 | 7.463 | |
| CaCl2 x 2 H2O | 0.05 | g | 0.05 | 0.339 | |
| Trace element solution SL-10 | 1.00 | ml | - | - | |
|
Brain heart infusion
(BD Bacto) |
6.00 | g | 5.988 | - | |
|
Proteose peptone
(BD Difco) |
6.00 | g | 5.988 | - | |
|
Yeast extract
(OXOID) |
2.00 | g | 1.996 | - | |
| Na-acetate | 1.00 | g | 0.998 | 12.166 | |
| Na-formate | 2.00 | g | 1.996 | 29.35 | |
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | 0.002 | |
| NaHCO3 | 4.00 | g | 3.992 | 47.52 | |
| Seven vitamins solution | 1.00 | ml | - | - | |
| L-Cysteine HCl x H2O | 0.50 | g | 0.499 | 2.841 | |
| Na2S x 9 H2O | 0.50 | g | 0.499 | 2.078 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Dissolve ingredients except bicarbonate, vitamins, cysteine, and sulfide, then 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. Add vitamins, cysteine, and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The vitamin solution should be sterilized by filtration. Before use, check the complete medium's pH and adjust it to 6.8 - 7.0, if necessary. | |||||
| 2 After growth has started and the culture is becoming turbid add sterile 80% H2 and 20% CO2 gas mixture to 0.5 - 1 bar overpressure. | |||||
Solution occurs in 265
other media (first defined in Medium 320).
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
|
HCl
(25%) |
10.0 | ml | 2.5 | 68.568 | |
| FeCl2 x 4 H2O | 1.5 | g | 1.5 | 7.545 | |
| ZnCl2 | 70.0 | mg | 0.07 | 0.514 | |
| MnCl2 x 4 H2O | 100.0 | mg | 0.1 | 0.505 | |
| H3BO3 | 6.0 | mg | 0.006 | 0.097 | |
| CoCl2 x 6 H2O | 190.0 | mg | 0.19 | 1.463 | |
| CuCl2 x 2 H2O | 2.0 | mg | 0.002 | 0.012 | |
| NiCl2 x 6 H2O | 24.0 | mg | 0.024 | 0.185 | |
| Na2MoO4 x 2 H2O | 36.0 | mg | 0.036 | 0.149 | |
| Distilled water | 990.0 | ml | - | - | |
| 1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. | |||||
Solution occurs in 78
other media (first defined in Medium 503).
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
|---|---|---|---|---|
| Vitamin B12 | 100 | mg | 0.1 | 0.074 |
| p-Aminobenzoic acid | 80 | mg | 0.08 | 0.583 |
| D-(+)-biotin | 20 | mg | 0.02 | 0.082 |
| Nicotinic acid | 200 | mg | 0.2 | 1.625 |
| Calcium pantothenate | 100 | mg | 0.1 | 0.21 |
| Pyridoxine hydrochloride | 300 | mg | 0.3 | 1.459 |
| Thiamine-HCl x 2 H2O | 200 | mg | 0.2 | 0.536 |
| Distilled water | 1000 | ml | - | - |
Metadata
| Last modified: | 28.02.23 |
| Source: | DSMZ |
| Taxonomic range: | Archaea |
| Medium type: | Complex medium |
| Final pH: | 6.8 - 7.0 |
| 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] |
|---|---|---|
| Brain heart infusion | 5.988 | - |
| Proteose peptone | 5.988 | - |
| NaHCO3 | 3.992 | 47.52 |
| Na-formate | 1.996 | 29.35 |
| Yeast extract | 1.996 | - |
| Show more | ||
Cultivation metadata from 35 strains
| pH | 4 - 6 | 1 strain |
| pH | 6 - 8 | 1 strain |
| pH | 8 - 10 | 1 strain |
| phyla | 1 | 33 strains |
| oxygen | anaerobe | 32 strains |
| Show more | ||