DSMZ 213 . SYNTHROPHOMONAS MEDIUM (SULFATE FREE)

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

Culture medium recipe

Vol.
×
Equipment needed: Gassing station Hungate tubes Autoclave Filter Centrifuge
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 952 ml - -
Solution B 30 ml - -
Solution C 1 ml - -
Solution D 10 ml - -
Solution E 10 ml - -
Solution F 10 ml - -
1 Add and dissolve ingredients of solution A, adjust pH to 7.2 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. Solution C is prepared under 100% N2 gas and sterilized by filtration. Solutions D, E and F are autoclaved under 100% N2 gas atmosphere. To complete the medium appropriate amounts of solutions B to F are added to the sterile solution A in in the sequence as indicated.
2 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.
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.525 3.859
MgCl2 x 6 H2O 0.33 g 0.347 1.705
NaCl 0.40 g 0.42 7.19
NH4Cl 0.40 g 0.42 7.855
CaCl2 x 2 H2O 0.05 g 0.053 0.357
Trace element solution SL-10 1.00 ml - -
Clarified rumen fluid 50.00 ml - -
Trypticase peptone
(BD BBL)
1.00 g 1.05 -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.3e-4 0.002
Distilled water 900.00 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2CO3 1.5 g 50 471.752
Distilled water 30.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Seven vitamins solution 1 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
Butyric acid 1.7 g 170 1929.41
Distilled water 10.0 ml - -
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Cysteine HCl x H2O 0.3 g 30 170.808
Distilled water 10.0 ml - -
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 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.
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: 28.02.22
Source: DSMZ
Taxonomic range:  Bacteria
Medium type: Complex medium
Final pH: 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: 6
Strains with modifications: 5

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Butyric acid 1.678 19.047
Na2CO3 1.481 13.971
Trypticase peptone 0.987 -
KH2PO4 0.494 3.627
NH4Cl 0.395 7.382
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Cultivation metadata from 6 strains BacDive

pH 6 - 8 2 strains
pH 8 - 10 2 strains
phyla 2 6 strains
oxygen anaerobe 6 strains
temperature mesophilic 6 strains