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DSMZ

DSMZ 1596 . METHANOCALCULUS ALKALIPHILUS MEDIUM

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

Culture medium recipe

Vol.
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Medium for Methanocalculus alkaliphilus DSM 24457

This strain uses the standard medium without modifications.

Growth conditions: 2-3 days anaerobic 30 °C

Main sol. 1596 #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 3.00 g 2.991 51.181
K2HPO4 1.00 g 0.997 5.724
NaHCO3 20.00 g 19.94 237.363
Na2CO3 15.00 g 14.955 141.102
NH4Cl 0.20 g 0.199 3.728
MgSO4 x 7 H2O 0.50 g 0.499 2.023
Trace elements solution (Pfennig & Lippert,1966) 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Na-formate 3.40 g 3.39 49.845
Na-acetate 0.16 g 0.16 1.945
Yeast extract
(OXOID) 		0.02 g 0.02 -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.25 g 0.249 1.038
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.

Trace elements solution (Pfennig & Lippert,1966) #

1000
Solution occurs in 28 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

Selenite-tungstate solution #

1000
Solution occurs in 179 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 - -

Wolin's vitamin solution (10x) #

1000
Solution occurs in 306 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: 02.01.23
Source: DSMZ
Taxonomic range:  Archaea
Taxonomic range:  Bacteria
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: 4
Strains with modifications: 1
Methanocalculus alkaliphilus DSM 24457

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 19.94 237.363
Na2CO3 14.955 141.102
Na-formate 3.39 49.845
NaCl 2.991 51.181
K2HPO4 0.997 5.724
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Cultivation metadata from 4 strains BacDive

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