DSMZ 892 . METHANOCALCULUS PUMILUS MEDIUM

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

Culture medium recipe

Vol.
×
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.40 g 0.392 2.882
NH4Cl 0.90 g 0.882 16.496
MgCl2 x 6 H2O 0.36 g 0.353 1.736
NaCl 10.00 g 9.804 167.76
Na-acetate 0.50 g 0.49 5.976
Yeast extract
(OXOID)
2.00 g 1.961 -
Trypticase peptone
(BD BBL)
2.00 g 1.961 -
Trace element solution 10.00 ml - -
Sodium resazurin
(0.1% w/v)
0.50 ml 4.9e-4 0.002
Na2CO3 5.00 g 4.902 46.25
Wolin's vitamin solution 10.00 ml - -
Na2S x 9 H2O 0.50 g 0.49 2.041
L-Cysteine HCl x H2O 0.50 g 0.49 2.791
Distilled water 1000.00 ml - -
1 Dissolve ingredients except carbonate, vitamins, cysteine and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve carbonate, adjust pH to 7.2 then dispense medium under 80% H2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add vitamins (sterilized by filtration), cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium and adjust to 7.3 - 7.5, if necessary. After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 1.5 - 2 bar overpressure.
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: 08.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.3 - 7.5
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 4.902 46.25
Trypticase peptone 1.961 -
Yeast extract 1.961 -
NH4Cl 0.882 16.496
Na-acetate 0.49 5.976
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Cultivation metadata from 1 strains BacDive

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