DSMZ 1257 . METHANOCELLA MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Hungate tubes Filter Autoclave Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaCl 1.0 g 1 17.112
MgCl2 x 6 H2O 0.4 g 0.4 1.968
CaCl2 x 2 H2O 0.1 g 0.1 0.68
NH4Cl 0.1 g 0.1 1.87
KH2PO4 0.2 g 0.2 1.47
KCl 0.5 g 0.5 6.707
Trace element solution 1.0 ml - -
Yeast extract
(OXOID)
0.1 g 0.1 -
Na-acetate 0.1 g 0.1 1.219
Sodium resazurin
(0.1% w/v)
0.5 ml 5.0e-4 -
Na2CO3 1.5 g 1.5 14.153
Wolin's vitamin solution 10.0 ml - -
L-Cysteine HCl x H2O 0.3 g 0.3 1.708
Na2S x 9 H2O 0.3 g 0.3 1.249
Distilled water 1000.0 ml - -
1 Dissolve ingredients (except carbonate, vitamins and reducing agents), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic and adjust pH to 6.0. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add vitamins, cysteine and sulfide from sterile anoxic stock solution prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Vitamins are sterilized by filtration. If necessary, adjust the pH of the complete medium to 7.0. After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.
Solution occurs in 14 other media (first defined in Medium 318).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid
(NTA)
12.80 g 12.8 66.967
FeCl2 x 4 H2O 1.00 g 1 5.03
MnCl2 x 4 H2O 0.10 g 0.1 0.505
CoCl2 x 6 H2O 0.03 g 0.03 0.231
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnCl2 0.10 g 0.1 0.734
CuCl2 0.02 g 0.02 0.149
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.03 g 0.03 0.124
NiCl2 x 6 H2O 0.10 g 0.1 0.772
NaCl 1.00 g 1 17.112
Na2SeO3 x 5 H2O 0.03 g 0.03 0.114
Na2WO4 x 2 H2O 0.04 g 0.04 0.121
Distilled water 1000.00 ml - -
1 First dissolve NTA in 200 ml of distilled water and adjust pH to 6.5 with KOH, then dissolve mineral salts. Finally adjust pH to 6.5 with KOH and 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: 03.05.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.0
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: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2CO3 1.5 14.153
KCl 0.5 6.707
MgCl2 x 6 H2O 0.4 1.968
L-Cysteine HCl x H2O 0.3 1.708
Na2S x 9 H2O 0.3 1.249
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Cultivation metadata from 1 strains BacDive

pH 6 - 8 1 strain
phyla 1 1 strain
oxygen anaerobe 1 strain
temperature thermophilic 1 strain