DSMZ 404 . METHANOSPHAERA CUNICULI MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Mineral solution 1 40.0 ml - -
Minerals solution 40.0 ml - -
Fe(NH4)2(SO4)2 x 7 H2O
(0.1% w/v)
2.0 ml 0.002 0.005
NiCl2 x 6 H2O
(0.1% w/v)
0.2 ml 2.0e-4 0.002
Modified Wolin's mineral solution 10.0 ml - -
Na-acetate x 3 H2O 3.0 g 3 -
Yeast extract
(OXOID)
1.0 g 1 -
Trypticase peptone
(BD BBL)
1.0 g 1 -
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 -
NaHCO3 3.0 g 3 35.711
Methanol 5.0 ml 3.96 123.588
Wolin's vitamin solution 10.0 ml - -
L-Cysteine HCl x H2O 0.3 g 0.3 1.708
Na2S x 9 H2O 0.3 g 0.3 1.249
Distilled water 900.0 ml - -
1 Dissolve ingredients (except bicarbonate, methanol, 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 7.0, then distribute under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add methanol, vitamins, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. Vitamins are sterilized by filtration. Adjust pH of complete medium to 7.0 - 7.2. After inoculation, pressurize culture bottles with sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure.
Solution occurs in 4 other media (first defined in Medium 203).
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 6 g 6 34.448
Distilled water 1000 ml - -
Solution occurs in 2 other media (first defined in Medium 317).
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 6.00 g 6 44.09
(NH4)2SO4 6.00 g 6 45.406
NaCl 12.00 g 12 205.339
MgSO4 x 7 H2O 2.60 g 2.6 10.549
CaCl2 x 2 H2O 0.16 g 0.16 1.088
Distilled water 1000.00 ml - -
Solution occurs in 170 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 357 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 -
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: 28.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.0 - 7.2
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Methanol 3.96 123.588
NaHCO3 3 35.711
Na-acetate x 3 H2O 3 -
Trypticase peptone 1 -
Yeast extract 1 -
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain