DSMZ 329 . METHANOHALOPHILUS HALOPHILUS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.329 6.157
KH2PO4 0.33 g 0.329 2.42
KCl 0.33 g 0.329 4.418
K2SO4 0.01 g 0.01 0.057
NaCl 70.00 g 69.86 1195.42
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID)
0.05 g 0.05 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
NaHCO3 5.00 g 4.99 59.4
MgCl2 x 6 H2O 0.33 g 0.329 1.62
CaCl2 x 2 H2O 0.33 g 0.329 2.24
Trimethylamine-HCl 5.00 g 4.99 -
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.705
Na2S x 9 H2O 0.15 g 0.15 0.623
Distilled water 1000.00 ml - -
1 Dissolve ingredients except bicarbonate, magnesium chloride, calcium chloride, vitamins, trimethylamine, cysteine and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add bicarbonate and equilibrate pH to 6.9 - 7.0, then distribute under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add magnesium chloride, calcium chloride, vitamins (sterilized by filtration), trimethylamine, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use adjust pH of complete medium to 7.2, if necessary.
Solution occurs in 263 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.
Solution occurs in 299 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: 25.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.2
Equipment needed: Autoclave, Filter, 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 69.86 1195.42
NaHCO3 4.99 59.4
Trimethylamine-HCl 4.99 -
NH4Cl 0.329 6.157
KCl 0.329 4.418
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Cultivation metadata from 1 strains BacDive

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