DSMZ 1596 . METHANOCALCULUS ALKALIPHILUS 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]
NaCl 3.00 g 3 51.335
K2HPO4 1.00 g 1 5.741
NaHCO3 20.00 g 20 238.075
Na2CO3 15.00 g 15 141.525
NH4Cl 0.20 g 0.2 3.739
MgSO4 x 7 H2O 0.50 g 0.5 2.029
Trace elements solution (Pfennig & Lippert,1966) 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Na-formate 3.40 g 3.4 49.995
Na-acetate 0.16 g 0.16 1.95
Yeast extract
(OXOID)
0.02 g 0.02 -
Wolin's vitamin solution 10.00 ml - -
Na2S x 9 H2O 0.25 g 0.25 1.041
Distilled water 1000.00 ml - -
1 Dissolve sodium chloride and hydrogenphosphate, then sparge solution with 100% N2 gas for at least 30 - 45 min to make it anoxic. Add and dissolve hydrogencarbonate and carbonate while gassing the head space only, then dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After autoclaving add ammonium chloride, magnesium sulfate, trace elements, formate, acetate, yeast extract, vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Vitamins should be sterilized by filtration. The pH of the complete medium should be 9.5.
2 Note: Addition of 10 - 20 mg sodium dithionite per liter (e.g. from 5% (w/v) solution, freshly prepared under N2 and filter-sterilized) may stimulate growth at the beginning. For transfers use 5 - 10% inoculum.
Solution occurs in 27 other media (first defined in Medium 1369).
Compound Amount Unit Conc. [g/L] Conc. [mM]
EDTA 5.00 g 5 17.109
FeSO4 x 7 H2O 2.20 g 2.2 7.913
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
MnCl2 x 4 H2O 0.03 g 0.03 0.152
H3BO3 0.03 g 0.03 0.485
CoCl2 x 6 H2O 0.20 g 0.2 1.54
CuCl2 x 2 H2O 0.03 g 0.03 0.176
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
Distilled water 1000.00 ml - -
1 pH 3.0-4.0
Solution occurs in 174 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 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: 11.05.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 9.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 100 % N2

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]
NaHCO3 20 238.075
Na2CO3 15 141.525
Na-formate 3.4 49.995
NaCl 3 51.335
K2HPO4 1 5.741
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Cultivation metadata from 1 strains BacDive

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