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DSMZ

DSMZ 375 . METHANOHALOBIUM MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 375 #

ml
Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 250.00 g 250 4277.89
NH4Cl 0.33 g 0.33 6.169
KH2PO4 0.33 g 0.33 2.425
KCl 0.33 g 0.33 4.427
CaCl2 x 2 H2O 0.33 g 0.33 2.245
MgCl2 x 6 H2O 0.33 g 0.33 1.623
MgSO4 x 7 H2O 4.00 g 4 16.229
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 -
Na2CO3 1.50 g 1.5 14.153
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID) 		0.05 g 0.05 -
Trimethylamine-HCl 5.00 g 5 -
Na2S x 9 H2O 0.50 g 0.5 2.082
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, trace elements, yeast extract, trimethylamine and sulfide. Sparge medium with 80% N2 and 20% CO2 gas mixture for at least 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add trace elements, yeast extract, trimethylamine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.4 with an anoxic sterile solution of 5% w/v Na2CO3, if necessary.
2 Note: A white precipitate that forms after autoclaving of the mineral base will dissolve again after storage at room temperature for 2 - 3 days.

Trace element solution SL-10 #

1000
Solution occurs in 265 other media (first defined in Medium 320).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(25%) 		10.0 ml 2.5 68.568
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 990.0 ml - -
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Metadata

Last modified: 28.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.4
Equipment needed: Autoclave, Gassing station, Hungate tubes
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Gas composition

  • 80 % N2
  • 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]
NaCl 250 4277.89
Trimethylamine-HCl 5 -
MgSO4 x 7 H2O 4 16.229
Na2CO3 1.5 14.153
Na2S x 9 H2O 0.5 2.082
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature thermophilic 1 strain