J484
484 .
MODIFIED METHANOCALDOCOCCUS MEDIUM
Inofficial medium: This medium was provided by external sources and is possibly not curated by experts.
Culture medium recipe
Vol.
×
Main sol. J484 #
ml
Equipment needed: Filter
Autoclave
Anaerobic workbench
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| K2HPO4 | 0.14 | g | 0.137 | 0.784 | |
| CaCl2 x 2 H2O | 0.14 | g | 0.137 | 0.929 | |
| NH4Cl | 0.25 | g | 0.244 | 4.56 | |
| MgSO4 x 7 H2O | 3.40 | g | 3.317 | 13.458 | |
| MgCl2 x 6 H2O | 2.70 | g | 2.634 | 12.957 | |
| KCl | 0.33 | g | 0.322 | 4.319 | |
| NaCl | 30.00 | g | 29.268 | 500.826 | |
| Iron stock solution | 5.00 | ml | - | - | |
| Trace minerals solution | 10.00 | ml | - | - | |
| Trace vitamins | 10.00 | ml | - | - | |
| Na2S2O3 x 5 H2O | 1.00 | g | 0.976 | 3.931 | |
|
Yeast extract
(BD-Difco) |
2.00 | g | 1.951 | - | |
| 2-Mercaptoethanol | 0.40 | g | 0.39 | 4.995 | |
| Resazurin | 1.00 | mg | 9.8e-4 | 0.004 | |
| Distilled water | 1000.00 | ml | - | - | |
| 1 Mix ingredients except Na2S2O3 x 5H2O, yeast extract, Trace vitamins and 2-mercaptoethanol, and adjust pH to 6.0. Bring to a boil for 5-10 sec, then, cool down under a stream of H2-CO2 (80:20, v/v). Dispense the medium into suitable culture vessels under a stream of H2-CO2 (80:20, v/v), seal with butyl rubber stoppers and autoclave 115C for 40 min. Separately autoclave Na2S2O3 x 5H2O and 2-mercaptoethanol as 50 g/L solutions and yeast extract as 100 g/L solution under a N2 atmosphere. Filter-sterilize Trace vitamins and store under a N2 atmosphere. Prior to inoculation, aseptically and anaerobically add the Na2S2O3 x 5H2O, yeast extract, Trace vitamins and 2-mercaptoethanol solutions to the medium. Pressurize the inoculated bottles to 200 kPa H2-CO2 (80:20, v/v). | |||||
| Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
|---|---|---|---|---|---|
| Nitrilotriacetic acid | 1.500 | g | 1.5 | 7.848 | |
| MgSO4 x 7 H2O | 3.000 | g | 3 | 12.172 | |
| MnSO4 x n H2O | 0.500 | g | 0.5 | - | |
| NaCl | 1.000 | g | 1 | 17.112 | |
| FeSO4 x 7 H2O | 0.100 | g | 0.1 | 0.36 | |
| CoSO4 x 7 H2O | 0.100 | g | 0.1 | 0.645 | |
| CaCl2 x 2 H2O | 0.100 | g | 0.1 | 0.68 | |
| ZnSO4 x 7 H2O | 0.100 | g | 0.1 | 0.348 | |
| CuSO4 x 5 H2O | 0.010 | g | 0.01 | 0.04 | |
| AlK(SO4)2 | 0.010 | g | 0.01 | 0.039 | |
| H3BO3 | 0.010 | g | 0.01 | 0.162 | |
| Na2MoO4 x 2 H2O | 0.010 | g | 0.01 | 0.041 | |
| NiCl2 x 6 H2O | 0.025 | g | 0.025 | 0.193 | |
| Na2SeO3 x 5 H2O | 0.300 | mg | 3.0e-4 | 0.001 | |
| Distilled water | 1000.000 | ml | - | - | |
| 1 Dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH. Then proceed to add minerals. Adjust final pH to 7.0. | |||||
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. |
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 |
| Medium type: | Complex medium |
| Final pH: | 6.5 |
| Equipment needed: | Anaerobic workbench, Autoclave, Filter |
| Edit in Medium builder | |