DSMZ 397 . FERULATE MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Filter Hungate tubes Gassing station Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 882 ml - -
Solution B 30 ml - -
Solution D 1 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
Sodium ferulate 10 mM - 10
1 Add and dissolve ingredients of solution A and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under the same gas atmosphere into anoxic Hungate-type tubes and autoclave. Solution B is prepared under 80% N2 and 20% CO2 gas atmosphere and autoclaved. Solutions E and F are autoclaved under 100% N2 gas atmosphere.
2 Solution D is prepared under 100% N2 gas and sterilized by filtration. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in the sequence as indicated.
3 Note: Some cultures are shipped in semi-solid medium which stimulates growth at the beginning. For agar stabs 3.00 g/l agar are added to the complete medium from a sterile anoxic stock solution (2% w/v). Upon receipt add anoxically 1 - 2 ml of the recommended freshly prepared liquid medium to the agar tube and incubate for 3 - 5 days . After incubation transfer 0.5 ml of the resulting cell suspension in the liquid phase to tubes with liquid medium.
4 Add sodium ferulate to 10 mM final concentration from filter-sterilized anaerobic stock solution.
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.567 4.166
MgCl2 x 6 H2O 0.33 g 0.374 1.84
NaCl 0.40 g 0.454 7.76
NH4Cl 0.40 g 0.454 8.479
CaCl2 x 2 H2O 0.05 g 0.057 0.386
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Clarified rumen fluid , clarified 50.00 ml - -
Trypticase peptone
(BD BBL)
1.00 g 1.134 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.7e-4 0.002
Distilled water 830.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Solution occurs in 1 other media (first defined in Medium 212).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Seven vitamins solution 1 ml - -
Solution occurs in 1 other media (first defined in Medium 212).
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 occurs in 1 other media (first defined in Medium 212).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.3 g 30 124.905
Distilled water 10.0 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 176 other media (first defined in Medium 385).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH 0.5 g 0.5 12.501
Na2SeO3 x 5 H2O 3.0 mg 0.003 0.011
Na2WO4 x 2 H2O 4.0 mg 0.004 0.012
Distilled water 1000.0 ml - -
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 - -
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: 17.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: n.d.
Equipment needed: Autoclave, 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

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2CO3 1.608 15.169
Trypticase peptone 1.072 -
KH2PO4 0.536 3.938
NH4Cl 0.429 8.015
NaCl 0.429 7.336
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Cultivation metadata from 1 strains BacDive

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