DSMZ 120d . METHANOSARCINA MARINE 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 0.35 2.009
KH2PO4 0.23 g 0.23 1.69
NH4Cl 0.50 g 0.5 9.348
MgCl2 x 6 H2O 10.00 g 10 49.188
CaCl2 x 2 H2O 0.15 g 0.15 1.02
NaCl 23.00 g 23 393.566
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 - -
Sodium resazurin
(0.1% w/v)
0.50 ml 5.0e-4 -
Na2CO3 1.00 g 1 9.435
Wolin's vitamin solution 10.00 ml - -
Methanol 10.00 ml - -
L-Cysteine HCl x H2O 0.30 g 0.3 -
Na2S x 9 H2O 0.30 g 0.3 1.249
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except carbonate, vitamins, methanol, cysteine and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. 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. 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. Carbonate is prepared under 80% N2 and 20% CO2 gas mixture and autoclaved separately. Vitamins 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 6.8 - 7.0, if necessary.
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: 22.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined 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

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 23 393.566
MgCl2 x 6 H2O 10 49.188
Na2CO3 1 9.435
NH4Cl 0.5 9.348
K2HPO4 0.35 2.009
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Cultivation metadata from 1 strains BacDive

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