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DSMZ

DSMZ 991 . CALDISPHAERA MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 991 #

ml
Equipment needed: Filter Water bath Gassing station Serum bottle Hungate tubes Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
(NH4)2SO4 1.30 g 1.286 9.731
KH2PO4 0.28 g 0.277 2.035
MgSO4 x 7 H2O 0.25 g 0.247 1.003
CaCl2 x 2 H2O 0.07 g 0.069 0.471
FeCl3 x 6 H2O 0.02 g 0.02 0.073
Allen's trace element solution 10.00 ml - -
Trisodium citrate x 2 H2O 2.94 g 2.908 9.888
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Sulfur
(powdered) 		10.00 g 9.891 308.425
Yeast extract
(OXOID) 		0.50 g 0.495 -
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.495 2.059
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except sulfur, yeast extract, vitamins and sulfide), adjust pH to 3.5 with 10 N H2SO4 and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 100 ml bottles) containing already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 4.0 - 4.5, if necessary.
2 After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.

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.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 305 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 17.10.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 4.0 - 4.5
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes, 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

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sulfur 9.891 308.425
Trisodium citrate x 2 H2O 2.908 9.888
(NH4)2SO4 1.286 9.731
Na2S x 9 H2O 0.495 2.059
Yeast extract 0.495 -
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Cultivation metadata from 1 strains BacDive

pH 2 - 4 1 strain
pH 4 - 6 1 strain
phyla 1 1 strain
nutrition chemoorganotroph 1 strain
oxygen anaerobe 1 strain
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