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DSMZ

DSMZ 825 . METHANOBACTERIUM II MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 825 #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 0.989 18.492
K2HPO4 0.3 g 0.297 1.704
KH2PO4 0.3 g 0.297 2.181
MgCl2 x 6 H2O 0.2 g 0.198 0.973
CaCl2 x 2 H2O 0.1 g 0.099 0.673
KCl 0.1 g 0.099 1.327
NaCl 0.6 g 0.593 10.155
Modified Wolin's mineral solution 10.0 ml - -
Na-acetate 0.5 g 0.495 6.029
Yeast extract 1.0 g 0.989 -
Sodium resazurin
(0.1% w/v) 		0.5 ml 4.9e-4 0.002
NaHCO3 4.0 g 3.956 47.097
Wolin's vitamin solution (10x) 1.0 ml - -
L-Cysteine HCl x H2O 0.5 g 0.495 2.816
Na2S x 9 H2O 0.5 g 0.495 2.059
Distilled water 1000.0 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, 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. Add 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.
2 After inoculation pressurize cultivation vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.

Modified Wolin's mineral solution #

1000
Solution occurs in 168 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.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 307 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: 27.10.22
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 % 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: 18
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 3.956 47.097
NH4Cl 0.989 18.492
Yeast extract 0.989 -
Na-acetate 0.495 6.029
L-Cysteine HCl x H2O 0.495 2.816
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Cultivation metadata from 18 strains BacDive

pH 4 - 6 1 strain
pH 6 - 8 1 strain
pH 8 - 10 1 strain
phyla 1 18 strains
oxygen anaerobe 18 strains
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