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DSMZ

DSMZ 141b . METHANOCULLEUS SP. MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. 141b #

ml
Equipment needed: Filter Autoclave Hungate tubes Gassing station
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.34 g 0.336 4.502
MgCl2 x 6 H2O 4.00 g 3.949 19.423
MgSO4 x 7 H2O 3.45 g 3.406 13.818
NH4Cl 0.25 g 0.247 4.614
CaCl2 x 2 H2O 0.14 g 0.138 0.94
K2HPO4 0.14 g 0.138 0.793
NaCl 6.00 g 5.923 101.352
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 -
Na-acetate 1.00 g 0.987 12.034
Yeast extract
(OXOID) 		2.00 g 1.974 -
Trypticase peptone
(BD BBL) 		2.00 g 1.974 -
Sodium resazurin
(0.1% w/v) 		0.50 ml 4.9e-4 0.002
Wolin's vitamin solution (10x) 1.00 ml - -
NaHCO3 5.00 g 4.936 58.755
L-Cysteine HCl x H2O 0.50 g 0.494 2.81
Na2S x 9 H2O 0.50 g 0.494 2.055
Distilled water 1000.00 ml - -
1 Dissolve ingredients (except bicarbonate, vitamins, cysteine and sulfide), sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Add and dissolve bicarbonate and adjust pH to 7.0, 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. After sterilization add cysteine and sulfide from sterile anoxic stock solutions autoclaved under 100% N2 gas atmosphere. Vitamins are prepared under 100% N2 gas atmosphere and sterilized by filtration. Adjust pH of final medium to 6.8 - 7.0.
2 For incubation use sterile 80% H2 and 20% CO2 gas mixture at two atmospheres of pressure.
3 Note: If the medium is being used without overpressure then adjust pH with a small amount of sterile anoxic 1 N HCl, if necessary.

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: 05.02.24
Source: DSMZ
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.8 - 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: 6
Strains with modifications: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 4.936 58.755
MgCl2 x 6 H2O 3.949 19.423
Yeast extract 1.974 -
Trypticase peptone 1.974 -
Na-acetate 0.987 12.034
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Cultivation metadata from 6 strains BacDive

pH 4 - 6 2 strains
pH 6 - 8 2 strains
pH 8 - 10 2 strains
phyla 1 6 strains
oxygen anaerobe 6 strains
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