DSMZ 141 . METHANOGENIUM MEDIUM (H2/CO2)

Official DSMZ medium: This medium was manually curated by experts from the DSMZ.

Culture medium recipe

Vol.
×
* Modified for Methanobacterium petrolearium DSM 22353

Adjust pH to 6.5.

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.34 g 0.333 4.463
MgCl2 x 6 H2O 4.00 g 3.914 19.252
MgSO4 x 7 H2O 3.45 g 3.376 13.696
NH4Cl 0.25 g 0.245 4.573
CaCl2 x 2 H2O 0.14 g 0.137 0.932
K2HPO4 0.14 g 0.137 0.786
NaCl 18.00 g 17.613 301.378
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 0.978 11.928
Yeast extract
(OXOID)
2.00 g 1.957 -
Trypticase peptone
(BD BBL)
2.00 g 1.957 -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
NaHCO3 5.00 g 4.892 58.238
Wolin's vitamin solution 10.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.489 2.786
Na2S x 9 H2O 0.50 g 0.489 2.037
Distilled water 1000.00 ml - -
1 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 7.0, 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.
2 For incubation use sterile 80% H2 and 20% CO2 gas mixture at two atmospheres of pressure.
3 Note: If the medium is being used without overpressure then adjust pH with a small amount of sterile anoxic 1 N HCl, if necessary.
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 56 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: 17.10.22
Source: DSMZ
Taxonomic range:  Archaea
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: * 6.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % H2
  • 20 % CO2

Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .

Associated strains

All strains for this medium: 31
Strains with modifications: 12
Methanobacterium petrolearium DSM 22353

  6.5

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 4.892 58.238
MgCl2 x 6 H2O 3.914 19.252
Yeast extract 1.957 -
Trypticase peptone 1.957 -
Na-acetate 0.978 11.928
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Cultivation metadata from 31 strains BacDive

pH 6 - 8 6 strains
pH 4 - 6 5 strains
pH 8 - 10 4 strains
phyla 2 31 strains
oxygen anaerobe 30 strains
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