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JCM 231 . METHANOBACTERIUM THERMOAUTOTROPHICUM MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. J231 #

ml
Equipment needed: Autoclave Anaerobic workbench
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.30 g 0.3 2.204
(NH4)2SO4 1.50 g 1.5 11.352
NaCl 0.60 g 0.6 10.267
MgSO4 x 7 H2O 0.12 g 0.12 0.487
CaCl2 x 2 H2O 0.08 g 0.08 0.544
FeSO4 x 7 H2O 4.00 mg 0.004 0.014
K2HPO4 0.15 g 0.15 0.861
Na2CO3 4.00 g 4 37.74
Trace vitamins 10.00 ml - -
Trace element solution 10.00 ml - -
L-Cysteine HCl x H2O 0.50 g 0.5 -
Na2S x 9 H2O 0.50 g 0.5 2.082
Resazurin 1.00 mg 1.0e-3 0.004
Distilled water 980.00 ml - -
1 Mix ingredients, except cysteine x HCl and Na2S x 9H2O, and adjust pH to 7.2. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20). Dispense the medium into suitable culture vessels (e.g., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20) and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Separately autoclave cysteine x HCl and Na2S x 9H2O as 5% solutions, respectively, under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the cysteine x HCl and Na2S x 9H2O solutions. Pressurize the inoculated bottles to 100 kPa H2-CO2 (80:20).

Trace element solution #

Solution occurs in 2 other media (first defined in Medium J231).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Na2-EDTA 0.640 g - -
MgSO4 x 7 H2O 6.200 g - -
MnSO4 x n H2O 0.550 g - -
NaCl 1.000 g - -
FeSO4 x 7 H2O 0.100 g - -
CoCl2 x 6 H2O 0.170 g - -
CaCl2 x 2 H2O 0.130 g - -
ZnSO4 x 7 H2O 0.180 g - -
CuSO4 x 5 H2O 0.050 g - -
AlK(SO4)2 x 12 H2O 0.018 g - -
H3BO3 0.010 g - -
Na2MoO4 x 2 H2O 0.011 g - -
NiCl2 x 6 H2O 0.025 g - -
1 Dissolve EDTA x 2Na first in 500 ml of distilled water. Then add the remaining salts and adjust volume to 1000 ml.

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 - -

Metadata

Last modified: 22.02.22
Source: JCM
Taxonomic range:  Archaea
Medium type: Defined medium
Final pH: 7.2
Equipment needed: Anaerobic workbench, Autoclave
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Associated strains

All strains for this medium: 3

Molecular composition

Compound Conc. [g/L] Conc. [mM]
Na2CO3 4 37.74
(NH4)2SO4 1.5 11.352
NaCl 0.6 10.267
Na2S x 9 H2O 0.5 2.082
L-Cysteine HCl x H2O 0.5 -
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Cultivation metadata from 3 strains BacDive

phyla 1 3 strains
oxygen anaerobe 3 strains
temperature thermophilic 3 strains