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DSMZ

DSMZ 892 . METHANOCALCULUS PUMILUS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 892 #

ml
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.40 g 0.396 2.907
NH4Cl 0.90 g 0.89 16.643
MgCl2 x 6 H2O 0.36 g 0.356 1.751
NaCl 10.00 g 9.891 169.254
Na-acetate 0.50 g 0.495 6.029
Yeast extract
(OXOID) 		2.00 g 1.978 -
Trypticase peptone
(BD BBL) 		2.00 g 1.978 -
Trace element solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Na2CO3 5.00 g 4.946 46.662
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.495 2.059
L-Cysteine HCl x H2O 0.50 g 0.495 2.816
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 to 30% of their volume 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.
2 After inoculation add sterile 80% H2 and 20% CO2 gas mixture to 1.5 - 2 bar overpressure.

Trace element solution #

1000
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.

Wolin's vitamin solution (10x) #

1000
Solution occurs in 305 other media (first defined in Medium 120).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 20 mg 0.02 0.082
Folic acid 20 mg 0.02 0.045
Pyridoxine hydrochloride 100 mg 0.1 0.486
Thiamine HCl 50 mg 0.05 0.148
Riboflavin 50 mg 0.05 0.133
Nicotinic acid 50 mg 0.05 0.406
Calcium D-(+)-pantothenate 50 mg 0.05 0.21
Vitamin B12 1 mg 1.0e-3 7.4e-4
p-Aminobenzoic acid 50 mg 0.05 0.365
(DL)-alpha-Lipoic acid 50 mg 0.05 0.242
Distilled water 1000 ml - -

Metadata

Last modified: 27.10.22
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 4.946 46.662
Yeast extract 1.978 -
Trypticase peptone 1.978 -
NH4Cl 0.89 16.643
Na-acetate 0.495 6.029
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Cultivation metadata from 1 strains BacDive

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