DSMZ 825a . METHANOBACTERIUM II MEDIUM (N2/CO2)

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

Culture medium recipe

Vol.
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* Modified for Methanobacterium oryzae DSM 11106

Supplement medium after autoclaving with 1.00 g/l Trypticase peptone and 3.00 g/l Na-formate added from sterile anoxic stock solutions prepared under 100% N2 gas.

Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.987 18.455
K2HPO4 0.3 g 0.296 1.7
KH2PO4 0.3 g 0.296 2.176
MgCl2 x 6 H2O 0.2 g 0.197 0.971
CaCl2 x 2 H2O 0.1 g 0.099 0.671
KCl 0.1 g 0.099 1.324
NaCl 0.6 g 0.592 10.135
Modified Wolin's mineral solution 10.0 ml - -
Na-acetate 0.5 g 0.494 6.017
Yeast extract 1.0 g 0.987 -
Sodium resazurin
(0.1% w/v)
0.5 ml 4.9e-4 0.002
NaHCO3 4.0 g 3.949 47.004
Methanol 1.5 ml 1.173 36.601
Wolin's vitamin solution (10x) 1.0 ml - -
L-Cysteine HCl x H2O 0.5 g 0.494 2.81
Na2S x 9 H2O 0.5 g 0.494 2.055
Na-formate* 3.0 g - -
Trypticase peptone* 1.0 g - -
Distilled water 1000.0 ml - -
1 Dissolve ingredients except bicarbonate, methanol, vitamins, cysteine and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate, adjust pH to 7.0, then dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add methanol, vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 6.8 - 7.0, 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 299 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 06.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % N2
  • 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: 3
Strains with modifications: 2
Methanobacterium oryzae DSM 11106

  3 g Na-formate
  1 g Trypticase peptone

Molecular composition
* modified beta

Compound Conc. [g/L] Conc. [mM]
NaHCO3 3.949 47.004
Na-formate 2.962 43.547
Methanol 1.173 36.601
NH4Cl 0.987 18.455
Trypticase peptone 0.987 -
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Cultivation metadata from 3 strains BacDive

phyla 1 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 3 strains