DSMZ 784a . METHANOCALCULUS NATRONOPHILUS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.2 g 0.2 1.47
MgCl2 x 6 H2O 0.1 g 0.1 0.492
NH4Cl 1.0 g 1 18.695
KCl 0.2 g 0.2 2.683
NaCl 60.0 g 60 1026.69
Trace element solution SL-11 1.0 ml - -
Na-acetate 0.2 g 0.2 2.438
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 -
Na2CO3 68.0 g 68 641.582
NaHCO3 38.0 g 38 452.343
Wolin's vitamin solution 10.0 ml - -
Na2S x 9 H2O 1.0 g 1 4.164
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate, bicarbonate, vitamins and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve carbonates and sulfide while gassing the head space only. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add vitamins from an anoxic stock solution sterilized by filtration. Adjust pH of the complete medium to 9.0 - 9.5.
Solution occurs in 22 other media (first defined in Medium 722).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA x 2 H2O 5.2 g 5.2 15.467
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 1000.0 ml - -
1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 ml.
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: 06.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 9.0 - 9.5
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2CO3 68 641.582
NaCl 60 1026.69
NaHCO3 38 452.343
NH4Cl 1 18.695
Na2S x 9 H2O 1 4.164
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Cultivation metadata from 1 strains BacDive

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