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DSMZ

DSMZ 391 . METHANOCOCCUS VOLTAE MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 391 #

ml
Equipment needed: Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
Mineral solution 2xW 500.0 ml - -
Modified Wolin's mineral solution 10.0 ml - -
LIP-solution 50.0 ml - -
Na-acetate 1.0 g 1 12.19
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 -
TYC-solution 50.0 ml - -
Na2CO3 1.5 g 1.5 14.153
L-Cysteine HCl x H2O 0.5 g 0.5 2.847
Na2S x 9 H2O 0.5 g 0.5 2.082
Distilled water 400.0 ml - -
1 Dissolve ingredients except TYC solution, carbonate, cystein and sulfide. Sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add TYC solution, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.2, if necessary.

Mineral solution 2xW #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 40.00 g 40 684.463
MgCl2 x 6 H2O 5.60 g 5.6 27.545
MgSO4 x 7 H2O 0.70 g 0.7 2.84
KCl 0.68 g 0.68 9.121
NH4Cl 0.50 g 0.5 9.348
K2HPO4 0.28 g 0.28 1.608
CaCl2 x 2 H2O 0.28 g 0.28 1.905
Distilled water 1000.00 ml - -

LIP-solution #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
L-Leucine 5.0 g 5 38.119
L-Isoleucine 10.0 g 10 76.237
Pantothenate 0.1 g 0.1 0.456
Distilled water 1000.0 ml - -
1 May be stored frozen without sterilization.

TYC-solution #

1000
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
Casamino acids
(BD Bacto) 		100 g 100 -
Yeast extract
(OXOID) 		50 g 50 -
L-Tryptophan 1 g 1 4.897
Distilled water 1000 ml - -
1 Prepare and autoclave under 100% N2 gas.

Modified Wolin's mineral solution #

1000
Solution occurs in 168 other media (first defined in Medium 141).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.50 g 1.5 7.848
MgSO4 x 7 H2O 3.00 g 3 12.172
MnSO4 x H2O 0.50 g 0.5 2.958
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.18 g 0.18 1.161
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.18 g 0.18 0.626
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 x 12 H2O 0.02 g 0.02 0.042
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
NiCl2 x 6 H2O 0.03 g 0.03 0.231
Na2SeO3 x 5 H2O 0.30 mg 3.0e-4 0.001
Na2WO4 x 2 H2O 0.40 mg 4.0e-4 0.001
Distilled water 1000.00 ml - -
1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH.

Metadata

Last modified: 28.02.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.2
Equipment needed: Autoclave, 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: 3
Strains with modifications: 2

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Casamino acids 5 -
MgCl2 x 6 H2O 2.8 13.773
Yeast extract 2.5 -
Na2CO3 1.5 14.153
Na-acetate 1 12.19
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Cultivation metadata from 3 strains BacDive

phyla 1 3 strains
oxygen anaerobe 3 strains
temperature mesophilic 3 strains