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JCM 265 . METHANOGENIUM MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. J265 #

ml
Equipment needed: Autoclave Anaerobic workbench Hungate tubes
Compound Amount Unit Conc. [g/L] Conc. [mM]
KCl 0.355 g 0.348 4.669
MgCl2 x 6 H2O 4.000 g 3.922 19.289
MgSO4 x 7 H2O 3.450 g 3.382 13.723
NH4Cl 0.250 g 0.245 4.582
CaCl2 x 2 H2O 0.140 g 0.137 0.934
KH2PO4 0.140 g 0.137 1.009
NaCl 18.000 g 17.647 301.969
Wolfe's mineral solution 10.000 ml - -
Trace vitamins 10.000 ml - -
Fe(NH4)2(SO4)2 x 6 H2O 2.000 mg 0.002 -
NaHCO3 5.000 g 4.902 58.352
Sodium acetate 1.000 g 0.98 11.952
Yeast extract
(BD-Difco) 		2.000 g 1.961 -
Trypticase peptone
(BD-BBL) 		2.000 g 1.961 -
Resazurin 1.000 mg 9.8e-4 0.004
Distilled water 980.000 ml - -
1 Mix components thoroughly, except NaHCO3, and adjust pH to 6.5. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20, v/v) and add NaHCO3 to the medium. Dispense the medium into suitable culture vessels (e.g., 5 ml medium in Hungate tubes) under the same gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Aseptically and anaerobically add per liter the following solutions (autoclaved and stored under a N2 atmosphere):
L-Cysteine HCl x H2O
(5%) 		10.000 ml - -
Na2S x 9 H2O
(5%) 		10.000 ml - -
2 Pressurize the inoculated bottles to 200 kPa H2:CO2 (80:20, v/v).

Wolfe's mineral solution #

1000
Solution occurs in 7 other media (first defined in Medium J265).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Trace minerals 1000.00 ml - -
NiCl2 x 6 H2O 0.02 g 0.02 0.154
Na2SeO3 1.00 mg 1.0e-3 -
Na2WO4 x 2 H2O 0.01 g 0.01 0.03

Trace vitamins #

1000
Solution occurs in 220 other media (first defined in Medium J197).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Biotin 2.0 mg 0.002 0.008
Folic acid 2.0 mg 0.002 0.005
Pyridoxine hydrochloride 10.0 mg 0.01 0.049
Thiamine HCl 5.0 mg 0.005 0.015
Riboflavin 5.0 mg 0.005 0.013
Nicotinic acid 5.0 mg 0.005 0.041
Calcium pantothenate 5.0 mg 0.005 0.01
Vitamin B12 0.1 mg 1.0e-4 7.4e-5
p-Aminobenzoic acid 5.0 mg 0.005 0.036
Lipoic acid 5.0 mg 0.005 0.024
Distilled water 1000.0 ml - -

Trace minerals #

1000
Solution occurs in 97 other media (first defined in Medium J151).
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 n H2O 0.50 g 0.5 -
NaCl 1.00 g 1 17.112
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.10 g 0.1 0.645
CaCl2 x 2 H2O 0.10 g 0.1 0.68
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
CuSO4 x 5 H2O 0.01 g 0.01 0.04
AlK(SO4)2 0.01 g 0.01 0.039
H3BO3 0.01 g 0.01 0.162
Na2MoO4 x 2 H2O 0.01 g 0.01 0.041
Distilled water 1000.00 ml - -
1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH solution. Then proceed to add minerals. Adjust final pH to 7.0.

Metadata

Last modified: 22.02.22
Source: JCM
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 6.5
Equipment needed: Anaerobic workbench, Autoclave, 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

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 17.657 302.137
NaHCO3 4.902 58.352
MgCl2 x 6 H2O 3.922 19.289
MgSO4 x 7 H2O 3.412 13.842
Yeast extract 1.961 -
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Cultivation metadata from 6 strains BacDive

phyla 1 6 strains
oxygen anaerobe 6 strains
temperature mesophilic 4 strains
temperature thermophilic 2 strains
temperature psychrophilic 1 strain