DSMZ 120c . METHANOSARCINA MAZEI (GOE1) MEDIUM

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

Culture medium recipe

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Medium for Methanosarcina mazei DSM 7222

This strain uses the standard medium without modifications.


Growth conditions: 3-7 days anaerobic 37 °C
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.35 g 0.349 2.001
KH2PO4 0.23 g 0.229 1.683
NH4Cl 0.50 g 0.498 9.31
MgSO4 x 7 H2O 0.50 g 0.498 2.021
CaCl2 x 2 H2O 0.25 g 0.249 1.694
NaCl 2.25 g 2.241 38.348
FeSO4 x 7 H2O solution (0.1% w/v) 2.00 ml - -
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID)
2.00 g 1.992 -
Casitone
(BD BBL)
2.00 g 1.992 -
Na-acetate 10.00 g 9.96 121.415
Sucrose 17.10 g 17.032 49.757
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 0.002
NaHCO3 2.00 g 1.992 23.713
Wolin's vitamin solution (10x) 1.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.299 1.701
Na2S x 9 H2O 0.30 g 0.299 1.244
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, vitamins, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then add and dissolve bicarbonate, adjust pH to 6.8 and dispense medium under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Cysteine and sulfide are autoclaved separately under 100% N2 gas atmosphere as concentrated stock solutions. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 6.8 - 7.0, if necessary.
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.
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.
Solution occurs in 306 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 22.02.22
Source: DSMZ
Synonyms: METHANOSARCINA MAZEI (GÖ1) MEDIUM
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 7.0
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 100 % 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
Methanosarcina mazei DSM 7222

No modifications necessary for this strain.

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Sucrose 17.032 49.757
Na-acetate 9.96 121.415
NaCl 2.241 38.348
NaHCO3 1.992 23.713
Casitone 1.992 -
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Cultivation metadata from 1 strains BacDive

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