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JCM 266 . METHANOMICROBIUM MOBILE MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. J266 #

ml
Equipment needed: Autoclave Anaerobic workbench
Compound Amount Unit Conc. [g/L] Conc. [mM]
Rumen fluid , clarified
(see below) 		300.00 ml - -
KH2PO4 0.30 g 0.3 2.204
K2HPO4 0.30 g 0.3 1.722
(NH4)2SO4 0.30 g 0.3 2.27
NaCl 0.60 g 0.6 10.267
MgSO4 x 7 H2O 0.13 g 0.13 0.527
CaCl2 x 2 H2O 8.00 mg 0.008 0.054
Trace minerals 10.00 ml - -
Trace vitamins 10.00 ml - -
FeSO4 x 7 H2O 2.00 mg 0.002 0.007
NaHCO3 2.00 g 2 23.808
Yeast extract
(BD-Difco) 		1.00 g 1 -
Trypticase peptone
(BD-BBL) 		1.00 g 1 -
Fatty acid mixture 20.00 ml - -
Resazurin 1.00 mg 1.0e-3 0.004
Na2S x 9 H2O 0.50 g 0.5 2.082
L-Cysteine HCl x H2O 0.50 g 0.5 -
Distilled water 660.00 ml - -
1 Mix ingredients, except NaHCO3, L-cysteine x HCl x H2O and Na2S x 9H2O, 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., 20 ml of the medium in 120 ml serum bottles) under a stream of H2-CO2 (80:20, v/v) 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 L-cysteine x HCl x H2O and Na2S x 9H2O solutions. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20, v/v).
2 To prepare the clarified rumen fluid, preheat rumen content at 120C for 15 min. Use the supernatant after centrifuging at 25,000 x g for 15 min.

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.

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.

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: Complex medium
Final pH: 7.5
Equipment needed: Anaerobic workbench, Autoclave
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaHCO3 2 23.808
Trypticase peptone 1 -
Yeast extract 1 -
NaCl 0.61 10.438
Isobutyric acid 0.5 5.675
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Cultivation metadata from 1 strains BacDive

phyla 1 1 strain
oxygen anaerobe 1 strain
temperature mesophilic 1 strain