DSMZ 760 . MARINE THERMOCOCCUS MEDIUM

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Culture medium recipe

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Medium for Thermococcus alcaliphilus DSM 10322

This strain uses the standard medium without modifications.


Growth conditions: 12-24 h anaerobic 85 °C pH 7.50
Equipment needed: Anaerobic workbench Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Medium 514 1000 ml - -
1 Prepare liquid Bacto Marine Broth (see medium 514) and filter through normal filter paper. An iron sediment will collect in the filter. Boil the medium in a water bath and cool under an anaerobic gas mixture (see strain dependant modifications).
2 Dispense the medium into Hungate tubes or serum bottles containing finely divided sulphur (0.5% w/v), seal the tubes or bottles under the same anaerobic gas used when cooling the medium. Sterilise the medium at 100°C for 3 hours on 3 consecutive days. Reduce the medium by adding 10% neutralised sodium sulphide to a final concentration of 0.05 %. The medium should not give a heavy black precipitate, if it does the iron sediment was not adequately removed by filtering in the initial stages and the medium should be made again, making sure that the iron is removed by filtering.
Solution occurs in 4 other media (first defined in Medium 514).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Bacto peptone 5.00 g 5 -
Yeast extract
(Bacto )
1.00 g 1 -
Fe(III) citrate 0.10 g 0.1 0.408
NaCl 19.45 g 19.45 332.82
MgCl2
(anhydrous)
5.90 g 5.9 61.968
Na2SO4 3.24 g 3.24 22.811
CaCl2 1.80 g 1.8 16.219
KCl 0.55 g 0.55 7.378
NaHCO3 0.16 g 0.16 1.905
KBr 0.08 g 0.08 0.672
SrCl2 34.00 mg 0.034 0.214
H3BO3 22.00 mg 0.022 0.356
Na-silicate 4.00 mg 0.004 0.033
NaF 2.40 mg 0.002 0.057
(NH4)NO3 1.60 mg 0.002 0.02
Na2HPO4 8.00 mg 0.008 0.056
Distilled water 1000.00 ml - -
1 Final pH should be 7.6 ± 0.2 at 25°C. If using the complete medium from Difco add 37.40 g to 1 litre water.

Metadata

Last modified: 19.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: n.d.
Equipment needed: Anaerobic workbench, Filter, 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: 32
Strains with modifications: 24
Thermococcus alcaliphilus DSM 10322

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 19.45 332.82
MgCl2 5.9 61.968
Bacto peptone 5 -
Na2SO4 3.24 22.811
CaCl2 1.8 16.219
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Cultivation metadata from 32 strains BacDive

phyla 1 33 strains
oxygen anaerobe 23 strains
temperature hyperthermophilic 29 strains
temperature thermophilic 2 strains