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DSMZ

DSMZ 491 . HYPERTHERMUS BUTYLICUS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 491 #

ml
Equipment needed: Hungate tubes Gassing station Water bath Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 17.00 g 16.553 283.249
MgSO4 x 7 H2O 3.50 g 3.408 13.827
MgCl2 x 6 H2O 2.75 g 2.678 13.171
KCl 0.33 g 0.321 4.31
NaBr 0.05 g 0.049 0.473
H3BO3 15.00 mg 0.015 0.236
SrCl2 x 6 H2O
(0.1% w/v) 		7.00 ml 0.007 0.026
(NH4)2SO4 10.00 mg 0.01 0.074
NH4Cl 0.50 g 0.487 9.102
Citric acid
(0.1% w/v) 		5.00 ml 0.005 0.025
KI
(0.1% w/v) 		2.50 ml 0.002 0.009
CaCl2 x 2 H2O 0.75 g 0.73 4.967
KH2PO4 0.50 g 0.487 3.578
NiCl2 x 6 H2O
(0.1% w/v) 		2.00 ml 0.002 0.015
Modified Wolin's mineral solution 10.00 ml - -
Tryptone
(BD BBL) 		6.00 g 5.842 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Sulfur
(powder) 		6.00 g 5.842 182.172
Na2S x 9 H2O 0.30 g 0.292 1.216
Distilled water 1000.00 ml - -
1 Dissolve ingredients except sulfur and sulfide and adjust pH to 6.0 with sulfuric acid. Sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic, dispense under the same gas atmosphere into anoxic Hungate-type tubes or serum vials containing the appropriate amount of sulfur, only to 30% of their volume to allow for a large headspace. Sterilize medium by heating for 2 - 3 hours in a boiling water bath on each of 3 successive days. Before use reduce the medium by adding sulfide from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. Check pH and adjust to pH 6.5 - 7.0, if necessary.
2 After inoculation pressurize culture vessels to 1 bar overpressure with a sterile 80% H2 and 20% CO2 gas mixture.

Modified Wolin's mineral solution #

1000
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: 27.10.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.5 - 7.0
Equipment needed: Autoclave, Gassing station, Hungate tubes, Water bath
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Gas composition

  • 50 % N2
  • 40 % H2
  • 10 % 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]
Sulfur 5.842 182.172
Tryptone 5.842 -
MgCl2 x 6 H2O 2.678 13.171
NH4Cl 0.487 9.102
KH2PO4 0.487 3.578
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Cultivation metadata from 1 strains BacDive

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