DSMZ 480d . THERMOPROTEUS UZONIENSIS MEDIUM

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Culture medium recipe

Vol.
×
Equipment needed: Anaerobic workbench Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Medium 480 1000 ml - -
Casitone 3 g/l 3 -
1 Use medium 480, but reduce the amount of Na acetate to 1.0 g/l and add 3.0 g/l casitone. Prepare sulphide and vitamin stock solutions anaerobically. Autoclave the sulphide solution but filter sterilise the vitamins. Prepare the medium anaerobically under N2:CO2 80:20. Adjust the pH to 5.7 with HCl. Sterilise the medium by heating for 3 hours on three successive days at 90-100°C. Add the vitamins and sulphide to the sterilised medium before use. The final pH should be 5.7-6.0. Dr. Bonch-Osmoloskaya supplied the information that the strain grows better on a peptone originally available in Russia. This product is no longer available, but it appears this product gave better growth than with highly purified products from Oxoid or Difco.
Solution occurs in 357 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 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 1 other media (first defined in Medium 480).
Equipment needed: Hungate tubes Filter Gassing station Autoclave Water bath
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.33 6.169
CaCl2 x 2 H2O 0.33 g 0.33 2.245
MgCl2 x 6 H2O 0.33 g 0.33 1.623
KCl 0.33 g 0.33 4.427
KH2PO4 0.33 g 0.33 2.425
Trace element solution SL-10 1.00 ml - -
Yeast extract 0.10 g 0.1 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Sulfur
(powder)
10.00 g 10 311.818
Na-acetate 5.00 g 5 60.951
NaHCO3 2.50 g 2.5 29.759
Wolin's vitamin solution 10.00 ml - -
Na2S x 9 H2O 0.50 g 0.5 2.082
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, acetate, bicarbonate, vitamins and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 5.9, dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur and sterilize by heating for 2-3 hours in a boiling water bath on each of 3 successive days. Add acetate, vitamins (sterilized by filtration)and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 6.8 - 7.0, if necessary.

Metadata

Last modified: 22.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 5.7 - 6.0
Equipment needed: Anaerobic workbench, Autoclave, Filter, Gassing station, Hungate tubes, Water bath
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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]
Sulfur 10 311.818
Na-acetate 5 60.951
Casitone 3 -
NaHCO3 2.5 29.759
Na2S x 9 H2O 0.5 2.082
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Cultivation metadata from 1 strains BacDive

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