88a .
SULFOLOBUS MEDIUM (ANAEROBIC)
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Equipment needed:
Autoclave
Gassing station
Hungate tubes
Water bath
Final gas composition:
100 % N2
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Final pH:
4.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
(NH4)2SO4 | 1.30 | g | ||
|
|
KH2PO4 | 0.28 | g | ||
|
|
MgSO4 x 7 H2O | 0.25 | g | ||
|
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CaCl2 x 2 H2O | 0.07 | g | ||
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FeCl3 x 6 H2O | 0.02 | g | ||
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|
Allen's trace element solution | 10.00 | ml | ||
|
|
Sulfur
(powder) |
10.00 | g | ||
|
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Yeast extract
(OXOID) |
0.50 | g | ||
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Na2S x 9 H2O | 0.50 | g | ||
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Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except sulfur, yeast extract and sulfide), adjust pH of the salt solution at room temperature to 4.0 using 1 N H2SO4 and sparge medium with 100% N2 gas to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials that contain already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a boiling water bath for 1 - 2 hours on each of 3 successive days. Add yeast extract and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to inoculation check pH and adjust to 4.0, if necessary. | |||||
Allen's trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MnCl2 x 4 H2O | 180 | mg | ||
|
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Na2B4O7 x 10 H2O | 450 | mg | ||
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ZnSO4 x 7 H2O | 22 | mg | ||
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CuCl2 x 2 H2O | 5 | mg | ||
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Na2MoO4 x 2 H2O | 3 | mg | ||
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VOSO4 x 2 H2O | 3 | mg | ||
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CoSO4 x 7 H2O | 1 | mg | ||
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Distilled water | 1000 | ml | ||
| 1 Adjust pH of final solution to 2 with 1 N HCl. | |||||