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DSMZ

DSMZ 784a . METHANOCALCULUS NATRONOPHILUS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 784a #

ml
Equipment needed: Autoclave Hungate tubes Filter Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.2 g 0.2 1.467
MgCl2 x 6 H2O 0.1 g 0.1 0.491
NH4Cl 1.0 g 0.998 18.658
KCl 0.2 g 0.2 2.677
NaCl 60.0 g 59.88 1024.64
Trace element solution SL-11 1.0 ml - -
Na-acetate 0.2 g 0.2 2.433
Sodium resazurin
(0.1% w/v) 		0.5 ml 5.0e-4 0.002
Na2CO3 68.0 g 67.864 640.301
NaHCO3 38.0 g 37.924 451.44
Wolin's vitamin solution (10x) 1.0 ml - -
Na2S x 9 H2O 1.0 g 0.998 4.155
Distilled water 1000.0 ml - -
1 Dissolve ingredients except carbonate, bicarbonate, vitamins and sulfide, then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve carbonates and sulfide while gassing the head space only. Dispense under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Before use add vitamins from an anoxic stock solution sterilized by filtration. Adjust pH of the complete medium to 9.0 - 9.5.

Trace element solution SL-11 #

1000
Solution occurs in 22 other media (first defined in Medium 722).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA x 2 H2O 5.2 g 5.2 15.467
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 1000.0 ml - -
1 Dissolve EDTA in 800 ml distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add other compounds. Finally adjust pH to 6.0 and bring volume to 1000 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: 06.04.22
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 9.0 - 9.5
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 67.864 640.301
NaCl 59.88 1024.64
NaHCO3 37.924 451.44
NH4Cl 0.998 18.658
Na2S x 9 H2O 0.998 4.155
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Cultivation metadata from 1 strains BacDive

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