Inofficial medium: This medium was provided by external sources and is possibly not curated by experts.
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. |
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. |
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. |
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 |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 7.5 |
Equipment needed: | Anaerobic workbench, Autoclave |
Edit in Medium builder |
All strains for this medium: | 1 |
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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