DSMZ 396 . METHANOSALSUM ZHILINAE MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract
(OXOID)
2.00 g 2 -
Trypticase peptone
(BD BBL)
2.00 g 2 -
NaCl 40.00 g 40 684.463
NH4Cl 1.00 g 1 18.695
MgCl2 x 6 H2O 3.50 g 3.5 17.216
MgSO4 x 7 H2O 3.00 g 3 12.172
KCl 1.00 g 1 13.414
K2HPO4 0.40 g 0.4 2.297
Modified Wolin's mineral solution 5.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
L-Cysteine HCl x H2O 0.50 g 0.5 2.847
NaHCO3 3.00 g 3 35.711
Na2CO3 2.00 g 2 18.87
Trimethylamine-HCl 2.00 g 2 -
Na2S x 9 H2O 0.25 g 0.25 1.041
Distilled water 1000.00 ml - -
1 Dissolve ingredients except cysteine, bicarbonate, carbonate, trimethylamine and sulfide, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add cysteine and dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add trimethylamine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas, whereas bicarbonate and carbonate are added from sterile anoxic stock solutions prepared under 80% N2 and 20% CO2 gas atmosphere. Adjust pH of complete medium to 9.2 - 9.4.
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: 28.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 9.2 - 9.4
Equipment needed: Autoclave, 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]
MgCl2 x 6 H2O 3.5 17.216
NaHCO3 3 35.711
Na2CO3 2 18.87
Trypticase peptone 2 -
Trimethylamine-HCl 2 -
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Cultivation metadata from 1 strains BacDive

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