897 .
IGNICOCCUS MEDIUM
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Equipment needed:
Gassing station
Hungate tubes
Water bath
Final pH:
5.0 - 5.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 13.65 | g | ||
|
|
MgSO4 x 7 H2O | 3.50 | g | ||
|
|
MgCl2 x 6 H2O | 2.75 | g | ||
|
|
KH2PO4 | 0.50 | g | ||
|
|
(NH4)2SO4 | 0.25 | g | ||
|
|
CaCl2 x 2 H2O | 0.38 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
NaBr | 0.05 | g | ||
|
|
H3BO3 | 15.00 | mg | ||
|
|
SrCl2 x 6 H2O
(0.1% w/v) |
7.00 | ml | ||
|
|
KI
(0.01% w/v) |
0.50 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Sulfur
(powdered) |
5.00 | g | ||
|
|
Meat extract | 1.00 | g | ||
|
|
Na2S x 9 H2O | 0.20 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except sulfur, meat extract and sulfide, adjust pH to 5.0 - 5.5 with 2 N H2SO4 and sparge medium with 80% H2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Distribute medium under the same gas atmosphere into anoxic Hungate-type tubes or serum vials already containing the appropriate amount of sulfur, only to 30% of their volume to allow for a large head space. For sterilization heat cultivation vessels in boiling water for 1 -2 hours on each of 3 successive days. Add meat extract and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere. | |||||
| 2 After inoculation pressurize the vessels with sterile 80% H2 and 20% CO2 gas mixture to 2 bar overpressure. | |||||