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JCM 228 . METHANOCOCCUS McC MEDIUM

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

Culture medium recipe

Vol.
×

Main sol. J228 #

ml
Equipment needed: Autoclave Anaerobic workbench
Compound Amount Unit Conc. [g/L] Conc. [mM]
General salts solution 500.00 ml - -
Trace minerals solution 10.00 ml - -
Iron stock solution 5.00 ml - -
NaCl 22.00 g 22 376.454
K2HPO4 0.14 g 0.14 0.804
Sodium acetate 1.36 g 1.36 16.579
Yeast extract
(BD-Difco) 		2.00 g 2 -
Resazurin 1.00 mg 1.0e-3 0.004
NaHCO3 5.00 g 5 59.519
L-Cysteine HCl x H2O 0.50 g 0.5 -
Na2S x 9 H2O 0.50 g 0.5 2.082
Distilled water 485.00 ml - -
1 Mix ingredients, except cysteine x HCl and Na2S x 9H2O, and bring to a boil for 5-10 sec. Cool down under a stream of N2-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), seal with butyl rubber stoppers, and autoclave. 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 vessels to 200 kPa H2-CO2 (80:20).

General salts solution #

Compound Amount Unit Conc. [g/L] Conc. [mM]
MgCl2 x 6 H2O 5.50 g - -
MgSO4 x 7 H2O 6.90 g - -
NH4Cl 1.00 g - -
KCl 0.67 g - -
CaCl2 x 2 H2O 0.28 g - -

Trace minerals solution #

1000
Solution occurs in 5 other media (first defined in Medium J228).
Compound Amount Unit Conc. [g/L] Conc. [mM]
Nitrilotriacetic acid 1.500 g 1.5 7.848
Fe(NH4)2(SO4)2 x 6 H2O 0.200 g 0.2 -
Na2SeO3 0.100 g 0.1 -
Na2MoO4 x 2 H2O 0.100 g 0.1 0.413
Na2WO4 x 2 H2O 1.000 g 1 3.032
MnSO4 x n H2O 0.100 g 0.1 -
ZnSO4 x 7 H2O 0.100 g 0.1 0.348
NiCl2 x 6 H2O 0.025 g 0.025 0.193
CuSO4 x 5 H2O 0.010 g 0.01 0.04
Distilled water 1000.000 ml - -
1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals.
2 Adjust final pH to 7.0.

Iron stock solution #

Solution occurs in 1 other media (first defined in Medium J228).
1 Dissolve 0.2 g of Fe(NH4)2(SO4)2 x 6H2O in 5 ml of distilled water containing 2 drops of concentrated HCl. When the Fe(NH4)2(SO4)2 x 6H2O crystal have dissolved, bring volume up to 100 ml.

Metadata

Last modified: 22.02.22
Source: JCM
Taxonomic range:  Archaea
Medium type: Complex medium
Final pH: 7.0
Equipment needed: Anaerobic workbench, Autoclave
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Associated strains

All strains for this medium: 4

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 22 376.454
NaHCO3 5 59.519
Yeast extract 2 -
Sodium acetate 1.36 16.579
Na2S x 9 H2O 0.5 2.082
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Cultivation metadata from 4 strains BacDive

phyla 1 4 strains
temperature mesophilic 4 strains