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JCM 872 . METHANOBACTERIUM MEDIUM (IV)

Inofficial medium: This medium was provided by external sources and is possibly not curated by experts.

Culture medium recipe

Vol.
×

Main sol. J872 #

ml
Equipment needed: Autoclave
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.50 g 0.489 3.595
MgSO4 x 7 H2O 0.40 g 0.391 1.588
NaCl 5.00 g 4.892 83.716
NH4Cl 0.40 g 0.391 7.317
CaCl2 x 2 H2O 0.05 g 0.049 0.333
FeSO4 x 7 H2O 2.00 mg 0.002 0.007
FeCl2 solution 1.00 ml - -
Trace element solution 1.00 ml - -
Yeast extract 1.00 g 0.978 -
Sludge fluid 50.00 ml - -
Sodium acetate 1.00 g 0.978 11.928
Sodium formate 2.00 g 1.957 28.776
NaHCO3 4.00 g 3.914 46.59
Fatty acid mixture 20.00 ml - -
Resazurin 1.00 mg 9.8e-4 0.004
Distilled water 930.00 ml - -
1 Mix components except NaHCO3 and bring to a boil for 5-10 sec. Cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3 to the medium. Dispense the medium into culture vessels under a stream of H2-CO2 (80:20, v/v), seal with butyl rubber stoppers and autoclave. Prior to inoculation, add per liter the following solutions (autoclaved and stored under a N2 atmosphere):
Na2S x 9 H2O
(5%) 		10.00 ml - -
L-Cysteine HCl x H2O
(5%) 		10.00 ml - -
2 Pressurize inoculated vessels to 200 kPa with a H2-CO2 (80:20, v/v) gas mixture.

Sludge fluid #

1000
Solution occurs in 7 other media (first defined in Medium 119).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Yeast extract 4 g 4 -
Sludge 1000 ml - -
1 Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with nitrogen gas for a few minutes incubate it at 37°C for 24 hours. Then centrifuge the sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped vessels under nitrogen gas. The sludge fluid can be stored at 8-12°C in the dark.

FeCl2 solution #

1000
Solution occurs in 87 other media (first defined in Medium J187).
Compound Amount Unit Conc. [g/L] Conc. [mM]
HCl
(7.7 M, 25%) 		10.0 ml 2.5 68.568
FeCl2 x 4 H2O 1.5 g 1.5 7.545
Distilled water 990.0 ml - -

Trace element solution #

1000
Solution occurs in 89 other media (first defined in Medium J187).
Compound Amount Unit Conc. [g/L] Conc. [mM]
ZnCl2 70 mg 0.07 0.514
MnCl2 x 4 H2O 100 mg 0.1 0.505
H3BO3 6 mg 0.006 0.097
CoCl2 x 6 H2O 190 mg 0.19 1.463
CuCl2 x 2 H2O 2 mg 0.002 0.012
NiCl2 x 6 H2O 24 mg 0.024 0.185
Na2MoO4 x 2 H2O 36 mg 0.036 0.149
Distilled water 1000 ml - -

Fatty acid mixture #

20
Solution occurs in 5 other media (first defined in Medium J266).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Valeric acid 0.5 g 25 244.786
Isovaleric acid 0.5 g 25 244.786
alpha-Methylbutyric acid 0.5 g 25 244.786
Isobutyric acid 0.5 g 25 283.736
Distilled water 20.0 ml - -
1 Adjust pH to 7.5 with concentrated NaOH solution.

Metadata

Last modified: 29.10.21
Source: JCM
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: n.d.
Equipment needed: Autoclave
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 4.892 83.716
NaHCO3 3.914 46.59
Sodium formate 1.957 28.776
Sodium acetate 0.978 11.928
Yeast extract 0.978 -
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature thermophilic 1 strain