DSMZ 120b . METHANOMICROCOCCUS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
K2HPO4 0.35 g 0.342 1.964
KH2PO4 0.23 g 0.225 1.652
NH4Cl 0.50 g 0.489 9.137
MgSO4 x 7 H2O 0.50 g 0.489 1.983
CaCl2 x 2 H2O 0.25 g 0.244 1.662
NaCl 2.25 g 2.199 37.635
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4)
2.00 ml 0.002 0.007
Trace element solution SL-10 1.00 ml - -
Yeast extract
(OXOID)
2.00 g 1.955 -
Casitone
(BD BBL)
2.00 g 1.955 -
Na-acetate 2.50 g 2.444 29.79
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
NaHCO3 2.50 g 2.444 29.09
2-Mercaptoethanesulfonate
(coenzyme M)
0.14 g 0.137 0.962
Wolin's vitamin solution 10.00 ml - -
Methanol 10.00 ml 7.742 241.618
L-Cysteine HCl x H2O 0.36 g 0.352 2.004
Na2S x 9 H2O 0.36 g 0.352 1.465
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, coenzyme M, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Then add and dissolve bicarbonate, adjust pH to 6.5 and dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Methanol (50% v/v stock solution) and the reducing agents are each autoclaved separately under 100% N2 gas atmosphere as concentrated solutions in tightly closed tubes. Vitamins and coenzyme M are prepared under 100% N2 gas atmosphere and sterilized by filtration. Appropriate volumes of the stock solutions are injected into the sterile medium with hypodermic syringes. Adjust pH of the complete medium to 7.0 - 7.2, if necessary. After inoculation, pressurize culture vessels with sterile 80% H2 and 20% CO2 gas mixture to 1 bar overpressure.
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 357 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium D-(+)-pantothenate 5.0 mg 0.005 -
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
(DL)-alpha-Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Metadata

Last modified: 23.08.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.0 - 7.2
Equipment needed: Autoclave, Filter, Gassing station, Hungate tubes
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Gas composition

  • 80 % H2
  • 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: 2
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Methanol 7.742 241.618
Na-acetate 2.444 29.79
NaHCO3 2.444 29.09
NaCl 2.199 37.635
Yeast extract 1.955 -
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Cultivation metadata from 2 strains BacDive

phyla 1 2 strains
oxygen anaerobe 2 strains
temperature mesophilic 2 strains