DSMZ 275 . TREPONEMA SUCCINIFACIENS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Gassing station Autoclave Hungate tubes Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 920 ml - -
Solution B 50 ml - -
Solution C 30 ml - -
1 Dissolve ingredients of solution A and sparge with 80% N2 and 20% CO2 gas atmosphere to make it anoxic. Then distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Solutions B is prepared under 100% N2 gas atmosphere and autoclaved. Solution C is prepared under 80% N2 and 20% CO2 gas atmosphere and autoclaved. To complete the medium appropriate amounts of solutions B and C are added to the sterile solution A in the sequence as indicated. Final pH of the medium should be at 7.4.
Compound Amount Unit Conc. [g/L] Conc. [mM]
CaCl2 x 2 H2O 0.1 g 0.109 0.739
MgSO4 x 7 H2O 0.1 g 0.109 0.441
KH2PO4 0.5 g 0.543 3.994
K2HPO4 0.5 g 0.543 3.12
NaCl 1.0 g 1.087 18.6
Clarified rumen fluid 300.0 ml - -
Yeast extract
(OXOID)
0.5 g 0.543 -
Peptone
(BD BACTO)
0.5 g 0.543 -
(NH4)2SO4 0.5 g 0.543 4.113
Sodium resazurin
(0.1% w/v)
0.5 ml 5.4e-4 0.002
L-Cysteine HCl x H2O 0.5 g 0.543 3.094
Distilled water 620.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Glucose 10 g 200 1110.12
Distilled water 50 ml - -
Equipment needed: Autoclave Hungate tubes
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Solution occurs in 27 other media (first defined in Medium 1310).
1 Rumen fluid from cow or sheep (obtained from fistulated animals or abattoir refuse) is filtered through muslin, autoclaved at 121°C for 15 min and then centrifuged at 27,000 g for 20 min. The supernatant is made anoxic by sparging with 100% N2 gas for 15 min, dispensed under same gas atmosphere into anoxic serum vials to 30% of volume and then stored frozen at -20°C.

Metadata

Last modified: 08.09.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.4
Equipment needed: Autoclave, Centrifuge, Gassing station, Hungate tubes
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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]
Glucose 10 55.506
Na2CO3 1.5 14.153
NaCl 1 17.112
(NH4)2SO4 0.5 3.784
KH2PO4 0.5 3.674
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain