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DSMZ

DSMZ 511 . METHANOPYRUS MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 511 #

ml
Equipment needed: Filter Autoclave Gassing station Serum bottle
Compound Amount Unit Conc. [g/L] Conc. [mM]
NH4Cl 0.25 g 0.248 4.641
K2HPO4 x 3 H2O 0.07 g 0.07 0.399
KH2PO4 0.09 g 0.089 0.657
NaCl 11.80 g 11.718 200.513
MgSO4 x 7 H2O 1.75 g 1.738 7.051
MgCl2 x 6 H2O 4.50 g 4.469 21.981
Na2SO4 0.81 g 0.804 5.663
CaCl2 x 2 H2O 0.78 g 0.775 5.269
KCl 0.30 g 0.298 3.996
Modified Wolin's mineral solution 10.00 ml - -
Fe(NH4)2(SO4)2 x 6 H2O
(0.1% w/v) 		2.00 ml 0.002 -
(NH4)2Ni(SO4)2
(0.1% w/v) 		2.00 ml 0.002 0.005
Na2WO4 x 2 H2O
(0.1% w/v) 		2.00 ml 0.002 0.006
Marine trace elements solution 10.00 ml - -
Sodium resazurin
(0.1% w/v) 		0.50 ml 5.0e-4 0.002
NaHCO3 1.00 g 0.993 11.821
Wolin's vitamin solution (10x) 1.00 ml - -
Na2S x 9 H2O 0.50 g 0.497 2.067
Distilled water 980.00 ml - -
1 Dissolve ingregients except bicarbonate, vitamins and sulfide, then 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 serum vials to 30% of their volume and autoclave. Add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Vitamins should be sterilized by filtration. Adjust pH of the complete medium to 6.5, if necessary.
2 After inoculation pressurize vessels to 2 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture.

Marine trace elements solution #

1000
Compound Amount Unit Conc. [g/L] Conc. [mM]
NaBr 4.00 g 4 38.877
SrCl2 x 6 H2O 1.80 g 1.8 6.751
H3BO3 1.30 g 1.3 21.022
KI 1.25 mg 0.001 0.005
Na-silicate 100.00 mg 0.1 0.819
NaF 60.00 mg 0.06 1.429
KNO3 40.00 mg 0.04 0.396
Na2HPO4 x 3 H2O 0.25 mg 2.5e-4 0.002
Distilled water 1000.00 ml - -

Modified Wolin's mineral solution #

1000
Solution occurs in 166 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.

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: Defined medium
Final pH: 6.5
Equipment needed: Autoclave, Filter, Gassing station, Serum bottle
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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]
MgCl2 x 6 H2O 4.469 21.981
NaHCO3 0.993 11.821
Na2SO4 0.804 5.663
Na2S x 9 H2O 0.497 2.067
KCl 0.298 3.996
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature hyperthermophilic 1 strain