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DSMZ

DSMZ 291 . PYROBACULUM NEUTROPHILUM MEDIUM

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

Culture medium recipe

Vol.
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Medium for Pyrobaculum neutrophilum DSM 2338

This strain uses the standard medium without modifications.

Growth conditions: 85 °C

Main sol. 291 #

ml
Equipment needed: Water bath Serum bottle Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.3 9.838
KH2PO4 0.28 g 0.28 2.058
MgSO4 x 7 H2O 0.25 g 0.25 1.014
CaCl2 x 2 H2O 0.07 g 0.07 0.476
FeCl3 x 6 H2O 0.02 g 0.02 0.074
Allen's trace element solution 10.00 ml - -
Distilled water 1000.00 ml - -
1 Prepare medium, then add per liter:
Sulfur
(powdered) 		8.00 g 8 249.454
Resazurin 0.40 mg 4.0e-4 0.002
neutralized Na2S x 9 H2O solution, 10% (w/v) 2.00 ml - -
2 Adjust pH to 6.5 with NaOH, bring to boil for 5 min, then cool to room temperature under H2 + CO2 (80% + 20%) gas mixture. Add 0.85 g NaHCO3 per liter and equilibrate the pH to c. 6.5 by further gassing.
3 Dispense in serum bottles (10 ml/30 ml bottle), seal and heat the bottles for one hour at 85°C on each of three successive days.
4 Before inoculation, add from anaerobic stock solutions 0.02% yeast extract (for mixotrophic growth only) and neutralized Na2S x 9 H2O solution, 10% (w/v). After inoculation, pressurize to 1 bar H2 + CO2 overpressure.

Allen's trace element solution #

1000
Solution occurs in 14 other media (first defined in Medium 88).
Compound Amount Unit Conc. [g/L] Conc. [mM]
MnCl2 x 4 H2O 180 mg 0.18 0.91
Na2B4O7 x 10 H2O 450 mg 0.45 -
ZnSO4 x 7 H2O 22 mg 0.022 0.077
CuCl2 x 2 H2O 5 mg 0.005 0.029
Na2MoO4 x 2 H2O 3 mg 0.003 0.012
VOSO4 x 2 H2O 3 mg 0.003 0.015
CoSO4 x 7 H2O 1 mg 1.0e-3 0.006
Distilled water 1000 ml - -
1 Adjust pH of final solution to 2 with 1 N HCl.

neutralized Na2S x 9 H2O solution, 10% (w/v) #

100
Solution occurs in 1 other media (first defined in Medium 573).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 10 g - -
Distilled water 100 ml - -
1 Adjust pH to 7.0.

Metadata

Last modified: 26.07.24
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 6.5
Equipment needed: Filter, Gassing station, Serum bottle, Water bath
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Gas composition

  • 80 % 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
Pyrobaculum neutrophilum DSM 2338

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sulfur 8 249.454
(NH4)2SO4 1.3 9.838
KH2PO4 0.28 2.058
MgSO4 x 7 H2O 0.25 1.014
CaCl2 x 2 H2O 0.07 0.476
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature hyperthermophilic 1 strain