Official DSMZ medium: This medium was manually curated by experts from the DSMZ.
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Solution A | 954 | ml | - | - | |
Solution B | 10 | ml | - | - | |
Solution C | 13 | ml | - | - | |
Solution D | 10 | ml | - | - | |
Solution E | 10 | ml | - | - | |
Solution F | 10 | ml | - | - | |
1 Dissolve ingredients of solution A, adjust pH to 5.0 with 0.1 N HCl and sparge with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense solution A under same gas atmosphere into anoxic Hungate-type tubes or serum vials (e.g., 20 ml medium in 50 ml bottles) and autoclave. Solutions B - F are prepared under 100% N2 gas atmosphere and sterilized by filtration. Prior to inoculation complete the medium by adding appropriate amounts of solutions B to F to the sterile solution A in the sequence as indicated. | |||||
2 Adjust pH of the complete medium to 5.1 with a sterile anoxic solution of 1 N HCl. After inoculation pressurize vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. | |||||
3 Note: Use 10-20% (v/v) as inoculum. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
NH4Cl | 27.0 | mg | 0.028 | 0.529 |
KH2PO4 | 14.0 | mg | 0.015 | 0.108 |
KCl
(0.1% w/v) |
2.0 | ml | 0.002 | 0.028 |
Na-acetate | 17.0 | mg | 0.018 | 0.217 |
Trace elements solution | 1.0 | ml | - | - |
Sodium resazurin
(0.1% w/v) |
0.5 | ml | 5.2e-4 | 0.002 |
Distilled water | 950.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
Homo-PIPES
(SIGMA) |
1.58 | g | 158 | - | |
NaOH
(pellets) |
0.18 | g | 18 | 450.034 | |
Distilled water | 10.00 | ml | - | - | |
1 Adjust pH of buffer to 5.5. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
NaOH
(0.38 M) |
7.20 | ml | 0.109 | 2.736 |
Nitrilotriacetate
(0.5 M, neutralized with 10 N NaOH) |
4.80 | ml | 0.459 | 2.4 |
TiCl3
(15% w/v in HCl, RIEDEL-de HAEN) |
0.55 | ml | 6.346 | 41.15 |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Yeast extract | 0.2 | g | 20 | - |
Distilled water | 10.0 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
2-Mercaptoethanesulfonate
(coenzyme M) |
0.07 | g | 7 | 49.226 |
Distilled water | 10.00 | ml | - | - |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] | |
---|---|---|---|---|---|
HCl
(25%) |
10.00 | ml | 2.475 | 67.889 | |
FeCl2 x 4 H2O | 1.34 | g | 1.327 | 6.673 | |
CoCl2 x 6 H2O | 24.00 | mg | 0.024 | 0.183 | |
ZnCl2 | 75.00 | mg | 0.074 | 0.545 | |
H3BO3 | 19.00 | mg | 0.019 | 0.304 | |
NiCl2 x 6 H2O | 24.00 | mg | 0.024 | 0.183 | |
Na2MoO4 x 2 H2O | 24.00 | mg | 0.024 | 0.098 | |
MnSO4 x 4 H2O | 26.00 | mg | 0.026 | 0.115 | |
MgSO4 x 7 H2O | 1.56 | g | 1.545 | 6.267 | |
CaCl2 x 2 H2O | 2.34 | g | 2.317 | 15.759 | |
CuSO4 x 5 H2O | 9.00 | mg | 0.009 | 0.036 | |
AlK(SO4)2 x 12 H2O | 3.45 | g | 3.416 | 7.201 | |
Distilled water | 1000.00 | ml | - | - | |
1 First dissolve ferrous chloride in the HCl, then dilute in water and dissolve the other salts. Finally make up to 1000 ml. |
Compound | Amount | Unit | Conc. [g/L] | Conc. [mM] |
---|---|---|---|---|
Biotin | 20 | mg | 0.02 | 0.082 |
Folic acid | 20 | mg | 0.02 | 0.045 |
Pyridoxine hydrochloride | 100 | mg | 0.1 | 0.486 |
Thiamine HCl | 50 | mg | 0.05 | 0.148 |
Riboflavin | 50 | mg | 0.05 | 0.133 |
Nicotinic acid | 50 | mg | 0.05 | 0.406 |
Calcium D-(+)-pantothenate | 50 | mg | 0.05 | 0.21 |
Vitamin B12 | 1 | mg | 1.0e-3 | 7.4e-4 |
p-Aminobenzoic acid | 50 | mg | 0.05 | 0.365 |
(DL)-alpha-Lipoic acid | 50 | mg | 0.05 | 0.242 |
Distilled water | 1000 | ml | - | - |
Last modified: | 03.05.22 |
Source: | DSMZ |
Taxonomic range: | Archaea |
Medium type: | Complex medium |
Final pH: | 5.1 |
Equipment needed: | Autoclave, Filter, Gassing station, Serum bottle |
Edit in Medium builder |
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] |
---|---|---|
Homo-PIPES | 1.569 | - |
Yeast extract | 0.199 | - |
TiCl3 | 0.082 | 0.531 |
2-Mercaptoethanesulfonate | 0.07 | 0.489 |
NH4Cl | 0.027 | 0.501 |
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