Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
NaCl | 20.0 | g | ||
|
NH4Cl | 1.0 | g | ||
|
K2HPO4 | 0.3 | g | ||
|
KH2PO4 | 0.3 | g | ||
|
KCl | 0.1 | g | ||
|
Na2SO4 | 2.0 | g | ||
|
CaCl2 x 2 H2O | 0.1 | g | ||
|
Trace element solution SL-10 | 1.0 | ml | ||
|
Yeast extract | 2.0 | g | ||
|
Sodium resazurin
(0.1% w/v) |
0.5 | ml | ||
|
Na2CO3 | 1.5 | g | ||
|
Na2S2O3 x 5 H2O | 3.0 | g | ||
|
MgCl2 x 6 H2O | 3.0 | g | ||
|
L-Cysteine HCl x H2O | 0.5 | g | ||
|
Na2S x 9 H2O | 0.5 | g | ||
|
Distilled water | 1000.0 | ml | ||
1 Dissolve ingredients (except carbonate, thiosulfate, magnesium chloride, cysteine and sulfide), then sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials to 30% of their volume and autoclave. Add magnesium chloride, cysteine and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of the complete medium to 6.8 - 7.0, if necessary. | |||||
2 After inoculation pressurize cultivation vials to 1 bar overpressure with sterile 80% H2 and 20% CO2 gas mixture. |
Compound | Amount | Unit | |||
---|---|---|---|---|---|
|
HCl
(25%) |
10.0 | ml | ||
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
ZnCl2 | 70.0 | mg | ||
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
H3BO3 | 6.0 | mg | ||
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
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. |