DSMZ 977 . VULCANITHERMUS MEDIUM

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Culture medium recipe

Vol.
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Equipment needed: Anaerobic workbench Hungate tubes Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.33 g 0.33 6.169
MgCl2 x 6 H2O 0.33 g 0.33 1.623
CaCl2 x 2 H2O 0.33 g 0.33 2.245
KCl 0.33 g 0.33 4.427
KNO3 0.33 g 0.33 3.264
NaCl 25.00 g 25 427.789
PIPES
(12mM)
3.60 g 3.6 11.906
Yeast extract 0.50 g 0.5 -
Tryptone 1.00 g 1 14.069
Sucrose 1.00 g 1 2.921
Modified Wolin's mineral solution II 1.00 ml - -
Wolin's vitamin solution 1.00 ml - -
Distilled water 1000.00 ml - -
1 Prepare the medium anaerobically, under nitrogen, omitting the CaCl2, MgCl2, KNO3, tryptone, yeast extract, vitamins and sucrose. The pH should be 6.8. Dispense the medium into vessels suitable for anaerobic growth (Hungate tubes or serum bottles) under an atmosphere of nitrogen and autoclave. To the sterile, cooled medium add, from sterile stock solutions the CaCl2, MgCl2, KNO3, tryptone, yeast extract, vitamins and sucrose. The CaCl2, MgCl2, KNO3, tryptone, yeast extract, and sucrose stock solutions should be autoclaved, while the vitamin solution is sterile filtered. This strain grows anaerobically in the presence of nitrate, but aerobic growth has been reported provided the oxygen concentration does not exceed 16% (v/v). Under anaerobic conditions a redox agent, such as sulphide is not required.
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 1 other media (first defined in Medium 534).
Equipment needed: Filter Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 30.00 g 30 513.347
MgSO4 x 7 H2O 7.00 g 7 28.4
MgCl2 x 6 H2O 5.50 g 5.5 27.053
KCl 0.65 g 0.65 8.719
NaBr 0.10 g 0.1 0.972
NH4Cl 0.15 g 0.15 2.804
K2HPO4 0.15 g 0.15 0.861
CaCl2 x 2 H2O 0.50 g 0.5 3.401
Trace element solution 10.00 ml - -
Distilled water 1000.00 ml - -
Solution A 30.00 ml - -
Solution B 30.00 ml - -
1 Gas with nitrogen gas for about 20 min and then adjust the pH to 6.5 - 6.8 with H2SO4. Fill 10 ml of medium in serum bottles (100 ml) under a protective CO2 atmosphere and tightly stopper and autoclave at 121°C for 15 min. Add 0,3 ml filter-sterilised solution A (gassed with N2) and sterile solution B (gassed with CO2) in 10 ml medium . Exchange the gas phase by a mixture of H2 : CO2 : O2 (300 KPa; each mixture 79.75 : 19.75 : 0.5 by volume). For more details see the original description (Huber et al. System. Appl. Microbiol. 15:340-351 (1992)).
Solution occurs in 1 other media (first defined in Medium 534).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Modified Wolin's mineral solution II 1000 ml - -
1 To 1000.0 ml trace element solution in medium 141 add:
(NH4)2Ni(SO4)2 2 g 2 5.063
Na2WO4 10 mg 0.01 0.034
Na2SeO4 10 mg 0.01 0.053
Solution occurs in 1 other media (first defined in Medium ).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaHCO3 2 g 66.667 793.585
Distilled water 30 ml - -
1 prepared under CO2 in an ice-bath, filter-sterilise
Solution occurs in 1 other media (first defined in Medium ).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S2O3 x 5 H2O 2 g 66.667 268.616
Distilled water 30 ml - -
1 Prepared under N2,autoclave 121°C, 15 min
Solution occurs in 22 other media (first defined in Medium 700).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.500 g - -
MgSO4 x 7 H2O 3.000 g - -
MnSO4 x H2O 0.500 g - -
NaCl 1.000 g - -
FeSO4 x 7 H2O 0.100 g - -
CoSO4 x 7 H2O 0.180 g - -
CaCl2 x 2 H2O 0.100 g - -
ZnSO4 x 7 H2O 0.180 g - -
CuSO4 x 5 H2O 0.010 g - -
AlK(SO4)2 x 12 H2O 0.020 g - -
H3BO3 0.010 g - -
Na2MoO4 x 2 H2O 0.010 g - -
NiCl2 x 6 H2O 0.025 g - -
Na2SeO3 x 5 H2O 0.300 mg - -
Distilled water 1000.000 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: 22.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 6.8
Equipment needed: Anaerobic workbench, Autoclave, Filter, Hungate tubes
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
PIPES 3.6 11.906
Tryptone 1 14.069
Sucrose 1 2.921
Yeast extract 0.5 -
NH4Cl 0.33 6.169
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen microaerophile 1 strain
temperature thermophilic 1 strain