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DSMZ

DSMZ 342 . METHANOBACTERIUM ALCALIPHILUM MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 342 #

ml
Equipment needed: Gassing station Hungate tubes Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 792 ml - -
Solution B 210 ml - -
Solution C 10 ml - -
1 Dissolve ingredients of solution A (except cysteine), bring to the boil, then cool to room temperature under 100% H2 gas. Add cysteine and adjust pH to 7.0, if necessary. Dispense medium under 100% H2 gas atmosphere into anoxic Hungate-type tubes or serum vials vials to 30% of their volume and then autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas atmosphere. Complete the medium by adding appropriate amounts of solutions B and C to the sterile solution A. Adjust pH of complete medium to 8.3 - 8.4, if necessary.
2 After inoculation add sterile H2 gas to 1 - 2 bar overpressure.

Solution A #

792
Equipment needed: Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.0 g 1.263 23.605
MgCl2 x 6 H2O 0.1 g 0.126 0.621
CaCl2 x 2 H2O 0.1 g 0.126 0.859
K2HPO4 0.4 g 0.505 2.9
Na2SeO3 x 5 H2O
(0.01% w/v) 		2.0 ml 2.5e-4 9.6e-4
Modified Wolin's mineral solution 10.0 ml - -
Yeast extract
(OXOID) 		2.0 g 2.525 -
Trypticase peptone
(BD BBL) 		2.0 g 2.525 -
Sodium resazurin
(0.1% w/v) 		0.5 ml 6.3e-4 0.003
L-Cysteine HCl x H2O 0.5 g 0.631 3.594
Distilled water 780.0 ml - -

Solution B #

210
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 10.0 g 47.619 566.846
Na2CO3 0.5 g 2.381 22.464
Distilled water 210.0 ml - -

Solution C #

10
Equipment needed: Hungate tubes Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Tris-HCl buffer
(pH 8.4, 2 M) 		10 ml - 20

Modified Wolin's mineral solution #

1000
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.

Metadata

Last modified: 27.10.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 8.3 - 8.4
Equipment needed: Autoclave, Gassing station, Hungate tubes
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Gas composition

  • 100 % H2

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

Associated strains

All strains for this medium: 4

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 9.881 117.626
Trypticase peptone 1.976 -
Yeast extract 1.976 -
NH4Cl 0.988 18.473
Na2CO3 0.494 4.662
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Cultivation metadata from 4 strains BacDive

phyla 1 4 strains
oxygen anaerobe 4 strains
temperature mesophilic 4 strains