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DSMZ

DSMZ 336 . OXOBACTER PFENNIGII MEDIUM

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

Culture medium recipe

Vol.
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Medium for Oxobacter pfennigii DSM 3222

This strain uses the standard medium without modifications.

Growth conditions: 2-3 days anaerobic 37 °C

Main sol. 336 #

ml
Equipment needed: Filter Gassing station Hungate tubes Autoclave Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 972 ml - -
Solution B 30 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
Solution E 3 ml - -
1 Sparge solution A with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, distribute under same gas atmosphere 8.9 ml medium per Hungate-type tube and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C is prepared under 100% N2 gas atmosphere and sterilized by filtration. Autoclave solutions D and E under 100% N2 gas. To complete the medium add appropriate amounts of the solutions B to E to the sterile solution A. Adjust pH of complete medium to 7.0 - 7.2, if necessary.

Solution A #

972
Compound Amount Unit Conc. [g/L] Conc. [mM]
Mineral solution 50.0 ml - -
Trace element solution SL-10 1.0 ml - -
Yeast extract 2.0 g 2.058 -
Na-vanillate 2.0 g 2.058 10.822
Clarified rumen fluid 300.0 ml - -
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 -
Distilled water 620.0 ml - -
1 Adjust pH to 6.9 before autoclaving.

Solution B #

30
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -

Solution C #

1
Compound Amount Unit Conc. [g/L] Conc. [mM]
Seven vitamins solution 1 ml - -

Solution D #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Cysteine HCl x H2O 0.3 g 30 170.808
Distilled water 10.0 ml - -

Solution E #

3
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 90 mg 30 124.905
Distilled water 3 ml - -

Mineral solution #

1000
Solution occurs in 4 other media (first defined in Medium 335).
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 10.0 g 10 73.484
MgCl2 x 6 H2O 6.6 g 6.6 32.464
NaCl 8.0 g 8 136.893
NH4Cl 8.0 g 8 149.561
CaCl2 x 2 H2O 1.0 g 1 6.802
Distilled water 1000.0 ml - -

Trace element solution SL-10 #

1000
Solution occurs in 265 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.

Clarified rumen fluid #

1000
Solution occurs in 28 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.

Seven vitamins solution #

1000
Solution occurs in 78 other media (first defined in Medium 503).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Vitamin B12 100 mg 0.1 0.074
p-Aminobenzoic acid 80 mg 0.08 0.583
D-(+)-biotin 20 mg 0.02 0.082
Nicotinic acid 200 mg 0.2 1.625
Calcium pantothenate 100 mg 0.1 0.21
Pyridoxine hydrochloride 300 mg 0.3 1.459
Thiamine-HCl x 2 H2O 200 mg 0.2 0.536
Distilled water 1000 ml - -

Metadata

Last modified: 25.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 7.0 - 7.2
Equipment needed: Autoclave, Centrifuge, Filter, 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
Oxobacter pfennigii DSM 3222

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na-vanillate 1.969 10.354
Yeast extract 1.969 -
Na2CO3 1.476 13.93
KH2PO4 0.492 3.616
NH4Cl 0.394 7.36
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Cultivation metadata from 1 strains BacDive

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