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DSMZ

DSMZ 905 . METHANOCALCULUS HALOTOLERANS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 905 #

ml
Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 1.00 g 0.99 18.51
K2HPO4 0.30 g 0.297 1.705
KH2PO4 0.30 g 0.297 2.183
KCl 0.17 g 0.168 2.258
NaCl 50.00 g 49.505 847.107
Na-acetate 0.50 g 0.495 6.035
Yeast extract
(OXOID) 		0.50 g 0.495 -
Trypticase
(BD BBL) 		0.50 g 0.495 -
Modified Wolin's mineral solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 2.00 g 1.98 23.572
L-Cysteine HCl x H2O 0.50 g 0.495 2.819
CaCl2 x 2 H2O 0.60 g 0.594 4.041
MgCl2 x 6 H2O 3.20 g 3.168 15.584
Na2S x 9 H2O 0.30 g 0.297 1.237
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, cysteine, calcium chloride, magnesium chloride and sulfide), then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve solid bicarbonate and cysteine, then adjust pH to 7.5 with 10 N KOH. Dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials (to 30% of volume) and autoclave. Add calcium chloride, magnesium chloride and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. The pH of the complete medium should be between 7.2 and 7.6.
2 After inoculation pressurize vessels with sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.

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: 7.2 - 7.6
Equipment needed: Autoclave, Gassing station, Hungate tubes
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Gas composition

  • 40 % N2
  • 40 % 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]
MgCl2 x 6 H2O 3.168 15.584
NaHCO3 1.98 23.572
NH4Cl 0.99 18.51
CaCl2 x 2 H2O 0.594 4.041
Na-acetate 0.495 6.035
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Cultivation metadata from 1 strains BacDive

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