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JCM 945 . MB MEDIUM FOR METHANOBACTERIUM

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

Culture medium recipe

Vol.
×

Main sol. J945 #

ml
Equipment needed: Autoclave Anaerobic workbench Filter
1 Mix components thoroughly, and autoclave under a N2-CO2 (80:20, v/v) gas atmosphere. After cooling, aseptically and anaerobically add the following solutions (filter-sterilized):
Compound Amount Unit Conc. [g/L] Conc. [mM]
KH2PO4 0.15 g 0.148 1.089
K2HPO4 0.30 g 0.296 1.702
NaCl 0.60 g 0.593 10.145
(NH4)2SO4 0.30 g 0.296 2.243
MgSO4 x 7 H2O 0.12 g 0.119 0.481
CaCl2 x 2 H2O 0.08 g 0.079 0.538
Trace minerals solution 10.00 ml - -
Resazurin 1.00 mg 9.9e-4 0.004
Distilled water 932.00 ml - -
NaHCO3
(8%) 		48.00 ml - -
Trace vitamins 10.00 ml - -
2 Distribute the medium into culture vessels (leave >80% gas phase by volume) under a H2-CO2 (80:20, v/v) gas mixture, seal with butyl rubber stoppers. Finally, add per liter the following solutions from anaerobic stocks (autoclaved and stored under a N2 atmosphere):
L-Cysteine HCl x H2O
(5%) 		6.00 ml - -
Na2S x 9 H2O
(5%) 		6.00 ml - -
3 Pressurize the inoculated vessels to 200 kPa H2-CO2 (80:20, v/v).

Trace minerals solution #

1000
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 -
FeSO4 x 7 H2O 0.10 g 0.1 0.36
CoSO4 x 7 H2O 0.10 g 0.1 0.645
ZnSO4 x 7 H2O 0.10 g 0.1 0.348
CuSO4 x 5 H2O 0.01 g 0.01 0.04
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. 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: Defined medium
Final pH: 6.5
Equipment needed: Anaerobic workbench, Autoclave, Filter
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Associated strains

All strains for this medium: 1

Molecular composition

Compound Conc. [g/L] Conc. [mM]
NaCl 0.593 10.145
(NH4)2SO4 0.296 2.243
K2HPO4 0.296 1.702
KH2PO4 0.148 1.089
MgSO4 x 7 H2O 0.148 0.601
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Cultivation metadata from 1 strains BacDive

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