DSMZ 1597a . METHANONATRONARCHAEUM MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Gassing station Filter Hungate tubes
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 500 ml - -
Solution B 500 ml - -
Solution C 1 ml - -
Solution D 1 ml - -
Solution E 5 ml - -
Solution F 2 ml - -
Solution G 10 ml - -
Solution H 5 ml - -
Solution I 10 ml - -
Solution J 10 ml - -
Solution K 5 ml - -
Solution L 10 ml - -
1 Sterilize solutions A and B in closed thick-walled screw-top bottles (e.g., SCHOTT) for 20 min at 120°C. The pH of solution A after sterilization should be 10. There is some precipitate forming that settles at the bottom after 3 - 4 days. It is best to remove precipitates by decantation before using solution A for medium preparation.
2 Combine solution A with solution B and sparge medium with 100% N2 gas 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. Solutions C to L are sterilized separately under 100% N2 gas. Vitamins and coenzyme M should be sterilized by filtration. To complete the medium appropriate amounts of solutions C to L are added to the combined sterile solutions A and B in the sequence as indicated. Final pH of the medium should be 9.5.
3 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.
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 92.5 g 185 1745.48
NaHCO3 17.5 g 35 416.632
NaCl 8.0 g 16 273.785
K2HPO4 0.5 g 1 5.741
Distilled water 500.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 120.00 g 240 4106.78
K2HPO4 1.25 g 2.5 14.353
KCl 2.50 g 5 67.069
Distilled water 500.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace elements solution (Pfennig & Lippert,1966) 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Selenite-tungstate solution 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.20 g 40 747.803
MgSO4 x 7 H2O 0.25 g 50 202.86
Distilled water 5.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Methanol 2 ml 792 24717.6
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na-formate 3.40 g 340 4999.47
Na-acetate 0.16 g 16 195.042
Distilled water 10.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 0.02 g 4 -
Distilled water 5.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
2-Mercaptoethanesulfonic acid
(coenzyme M)
0.15 g 15 105.485
Distilled water 10.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Wolin's vitamin solution 10 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Ferrous sulfide sludge 5 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.25 g 25 104.088
Distilled water 10.00 ml - -
Solution occurs in 27 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
Solution occurs in 173 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 - -
Solution occurs in 356 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -
Solution occurs in 3 other media (first defined in Medium 1267).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 15.4 g 154 553.927
Na2S x 9 H2O 12.3 g 123 512.112
Distilled water 100.0 ml - -
1 Heat distilled water to 50°C in a 250 ml beaker with a stir bar present. While rapidly stirring the water, add the ferrous sulfate followed immediately by the sodium sulfide. The formed black FeS sludge is decanted into a glass bottle that can be stoppered. The FeS is allowed to settle for several hours and then the overlying water is decanted and replaced. This procedure is repeated at least five times to wash the FeS. After washing, the pH of the FeS solution should be close to neutrality. The FeS suspension can be kept in closed bottles or tubes under a nitrogen atmosphere for at least three months.

Metadata

Last modified: 11.05.22
Source: DSMZ
Taxonomic range:  Archaea
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 113.314 1938.99
Na2CO3 87.347 824.117
NaHCO3 16.525 196.71
Na-formate 3.211 47.209
KCl 2.361 31.666
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Cultivation metadata from 1 strains BacDive

halophily extremely halophilic 1 strain
pH 8 - 10 1 strain
pH 10 - 12 1 strain
phyla 1 1 strain
nutrition heterotroph 1 strain
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