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DSMZ

DSMZ 213 . SYNTHROPHOMONAS MEDIUM (SULFATE FREE)

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

Culture medium recipe

Vol.
×

Main sol. 213 #

ml
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.

Solution A #

952
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 - -

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]
Butyric acid 1.7 g 170 1929.41
Distilled water 10.0 ml - -

Solution E #

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 F #

10
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2S x 9 H2O 0.3 g 30 124.905
Distilled water 10.0 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: 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