Inofficial medium: This medium was provided by external sources and is possibly not curated by experts.
1 Mix components thoroughly and adjust pH to 7.2. Distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a stream of N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. Allow to stand overnight. Aseptically and anaerobically add the following solutions (filter-sterilized): | |||||
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NaCl | 1.00 | g | 1.008 | 17.25 | |
MgCl2 x 6 H2O | 0.20 | g | 0.202 | 0.992 | |
NaH2PO4 x 2 H2O | 0.91 | g | 0.917 | 7.646 | |
CaCl2 x 2 H2O | 0.10 | g | 0.101 | 0.686 | |
NH4Cl | 1.00 | g | 1.008 | 18.846 | |
Yeast extract
(BD-Difco) |
0.20 | g | 0.202 | - | |
FeCl2 solution | 1.00 | ml | - | - | |
Trace element solution | 1.00 | ml | - | - | |
Distilled water | 900.00 | ml | - | - | |
NaHCO3
(8.0%) |
30.00 | ml | - | - | |
Trace vitamins | 10.00 | ml | - | - | |
Lactose
(0.25 M) |
50.00 | ml | - | - | |
2 Readjust pH to 7.2-7.6, if necessary. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
HCl
(7.7 M, 25%) |
10.0 | ml | 2.5 | 68.568 |
FeCl2 x 4 H2O | 1.5 | g | 1.5 | 7.545 |
Distilled water | 990.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
ZnCl2 | 70 | mg | 0.07 | 0.514 |
MnCl2 x 4 H2O | 100 | mg | 0.1 | 0.505 |
H3BO3 | 6 | mg | 0.006 | 0.097 |
CoCl2 x 6 H2O | 190 | mg | 0.19 | 1.463 |
CuCl2 x 2 H2O | 2 | mg | 0.002 | 0.012 |
NiCl2 x 6 H2O | 24 | mg | 0.024 | 0.185 |
Na2MoO4 x 2 H2O | 36 | mg | 0.036 | 0.149 |
Distilled water | 1000 | ml | - | - |
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 | - | - |
Last modified: | 22.02.22 |
Source: | JCM |
Medium type: | Complex medium |
Final pH: | 7.4 |
Equipment needed: | Anaerobic workbench, Autoclave, Filter, Hungate tubes |
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