This medium was extracted automatically and has not been curated by experts, yet.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Medium 480 | 1000 | ml | - | - | |
Casitone | 3 | g/l | 3 | - | |
1 Use medium 480, but reduce the amount of Na acetate to 1.0 g/l and add 3.0 g/l casitone. Prepare sulphide and vitamin stock solutions anaerobically. Autoclave the sulphide solution but filter sterilise the vitamins. Prepare the medium anaerobically under N2:CO2 80:20. Adjust the pH to 5.7 with HCl. Sterilise the medium by heating for 3 hours on three successive days at 90-100°C. Add the vitamins and sulphide to the sterilised medium before use. The final pH should be 5.7-6.0. Dr. Bonch-Osmoloskaya supplied the information that the strain grows better on a peptone originally available in Russia. This product is no longer available, but it appears this product gave better growth than with highly purified products from Oxoid or Difco. |
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 D-(+)-pantothenate | 5.0 | mg | 0.005 | 0.021 |
Vitamin B12 | 0.1 | mg | 1.0e-4 | 7.4e-5 |
p-Aminobenzoic acid | 5.0 | mg | 0.005 | 0.036 |
(DL)-alpha-Lipoic acid | 5.0 | mg | 0.005 | 0.024 |
Distilled water | 1000.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
HCl
(25%) |
10.0 | ml | 2.5 | 68.568 | |
FeCl2 x 4 H2O | 1.5 | g | 1.5 | 7.545 | |
ZnCl2 | 70.0 | mg | 0.07 | 0.514 | |
MnCl2 x 4 H2O | 100.0 | mg | 0.1 | 0.505 | |
H3BO3 | 6.0 | mg | 0.006 | 0.097 | |
CoCl2 x 6 H2O | 190.0 | mg | 0.19 | 1.463 | |
CuCl2 x 2 H2O | 2.0 | mg | 0.002 | 0.012 | |
NiCl2 x 6 H2O | 24.0 | mg | 0.024 | 0.185 | |
Na2MoO4 x 2 H2O | 36.0 | mg | 0.036 | 0.149 | |
Distilled water | 990.0 | ml | - | - | |
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
NH4Cl | 0.33 | g | 0.33 | 6.169 | |
CaCl2 x 2 H2O | 0.33 | g | 0.33 | 2.245 | |
MgCl2 x 6 H2O | 0.33 | g | 0.33 | 1.623 | |
KCl | 0.33 | g | 0.33 | 4.427 | |
KH2PO4 | 0.33 | g | 0.33 | 2.425 | |
Trace element solution SL-10 | 1.00 | ml | - | - | |
Yeast extract | 0.10 | g | 0.1 | - | |
Sodium resazurin
(0.1% w/v) |
0.50 | ml | 5.0e-4 | - | |
Sulfur
(powder) |
10.00 | g | 10 | 311.818 | |
Na-acetate | 5.00 | g | 5 | 60.951 | |
NaHCO3 | 2.50 | g | 2.5 | 29.759 | |
Wolin's vitamin solution | 10.00 | ml | - | - | |
Na2S x 9 H2O | 0.50 | g | 0.5 | 2.082 | |
Distilled water | 1000.00 | ml | - | - | |
1 Dissolve ingredients (except sulfur, acetate, bicarbonate, vitamins and sulfide) and sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Adjust pH to 5.9, dispense under 80% N2 and 20% CO2 gas atmosphere into anoxic Hungate-type tubes or serum vials containing already the appropriate amount of sulfur and sterilize by heating for 2-3 hours in a boiling water bath on each of 3 successive days. Add acetate, vitamins (sterilized by filtration)and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Prior to use adjust pH of complete medium to 6.8 - 7.0, if necessary. |
Last modified: | 22.02.22 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 5.7 - 6.0 |
Equipment needed: | Anaerobic workbench, Autoclave, Filter, Gassing station, Hungate tubes, Water bath |
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Detailed instructions for the cultivation of anaerobes including important warnings can be found in the cultivation instructions .
All strains for this medium: | 1 |
Compound | Conc. [g/L] | Conc. [mM] |
---|---|---|
Sulfur | 10 | 311.818 |
Na-acetate | 5 | 60.951 |
Casitone | 3 | - |
NaHCO3 | 2.5 | 29.759 |
Na2S x 9 H2O | 0.5 | 2.082 |
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