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DSMZ

DSMZ 503 . ANAEROBIC FRESHWATER (FWM) MEDIUM

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

Culture medium recipe

Vol.
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Medium for Methanomethylovorans uponensis DSM 27305

* Modification: Supplement medium with 20 ml/l FeSO4 x 7 H2O solution (0.1% w/v), 1 ml/l Na-dithionite solution (5% w/v), 0.8 ml/l Methanol, and 2 g/l Trimethylamine-HCl. Omit D-Glucose.

Growth conditions: 3-7 days anaerobic 35 °C

Main sol. 503 #

ml
Equipment needed: Autoclave Gassing station Hungate tubes Filter
Compound Amount Unit Conc. [g/L] Conc. [mM]
Solution A 942 ml - -
Solution B 30 ml - -
Solution C 20 ml - -
Solution D 1 ml - -
Solution E 10 ml - -
Na-dithionite solution (5% w/v)* 1 ml - -
FeSO4 x 7 H2O solution (0.1% w/v)* 20 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 into anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere. Solution C and D are prepared under 100% N2 gas and sterilized by filtration. Solution E is autoclaved under 100% N2 gas. To complete the medium appropriate amounts of solutions B to E are added to the sterile solution A in the sequence as indicated. Adjust pH of complete medium to 7.2 - 7.4, if necessary.

Solution A #

942
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.20 g 0.212 1.56
NH4Cl 0.25 g 0.265 4.962
NaCl 1.00 g 1.062 18.165
MgCl2 x 6 H2O 0.40 g 0.425 2.089
KCl 0.50 g 0.531 7.12
CaCl2 x 2 H2O 0.15 g 0.159 1.083
Trace element solution SL-10 1.00 ml - -
Selenite-tungstate solution 1.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.3e-4 0.002
Distilled water 940.00 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 #

20
Compound Amount Unit Conc. [g/L] Conc. [mM]
D-Glucose* 5.0 g 250 1387.66
Trimethylamine-HCl* 2.0 g - -
Methanol* 0.8 ml - -
Distilled water 20.0 ml - -

Solution D #

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

Solution E #

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 266 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.

Selenite-tungstate solution #

1000
Solution occurs in 179 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 - -

Seven vitamins solution #

1000
Solution occurs in 77 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 - -

Na-dithionite solution (5% w/v) * #

10
Solution occurs in 7 other media (first defined in Medium 829).
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaOH
(1 N) 		0.5 ml - -
Na2S2O4 0.5 g 50 287.175
Distilled water 9.5 ml - -
1 Add 1 N NaOH to distilled water and make the solution anoxic by sparging with 100% N2 gas. Then dissolve the Na-dithionite and filter-sterilize the solution into anoxic Hungate tubes. The final pH of the solution should be between 9 and 10. Store the prepared solution in a dark and cool place. Prepare only small amounts of stock solution, as Na-dithionite decomposes rapidly.

FeSO4 x 7 H2O solution (0.1% w/v) * #

1000
Solution occurs in 13 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
FeSO4 x 7 H2O 1 g 1 3.597
H2SO4
(0.1 N) 		1000 ml - -
1 The ferrous sulfate solution is not stable and should be freshly prepared.

Metadata

Last modified: 26.09.22
Source: DSMZ
Taxonomic range:  Archaea
Taxonomic range:  Bacteria
Medium type: Defined medium
Final pH: 7.2 - 7.4
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: 35
Strains with modifications: 35
Methanomethylovorans uponensis DSM 27305

+  1 ml Na-dithionite solution (5% w/v)
+  20 ml FeSO4 x 7 H2O solution (0.1% w/v)
  D-Glucose
  2 g Trimethylamine-HCl
  0.8 ml Methanol

Molecular composition

Compound Conc. [g/L] Conc. [mM]
D-Glucose 4.985 27.67
Na2CO3 1.496 14.11
NaCl 0.997 17.06
KCl 0.499 6.687
MgCl2 x 6 H2O 0.399 1.962
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Cultivation metadata from 35 strains BacDive

pH 6 - 8 7 strains
pH 8 - 10 3 strains
pH 4 - 6 1 strain
phyla 12 35 strains
nutrition chemoorganoheterotroph 1 strain
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