507a .
CARBOXYDOTHERMUS SIDEROPHILUS MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.9
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KCl | 0.33 | g | ||
|
|
MgCl2 x 6 H2O | 0.52 | g | ||
|
|
CaCl2 x 2 H2O | 0.29 | g | ||
|
|
NH4Cl | 0.33 | g | ||
|
|
KH2PO4 | 0.33 | g | ||
|
|
Na2-9,10-anthraquinone-2,6-disulfonate
(TCI A0308) |
8.25 | g | ||
|
|
Trace element solution SL-4 | 10.00 | ml | ||
|
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NaHCO3 | 1.00 | g | ||
|
|
Yeast extract | 0.20 | g | ||
|
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Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except bicarbonate, yeast extract, vitamins and sulfide, then sparge medium with 100% N2 gas 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 yeast extract, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas atmosphere and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. After completing the medium adjust pH to 6.8 - 7.0. | |||||
| 2 Inoculated vessels are pressurized with sterile carbon monoxide gas to 1 bar overpressure. | |||||
Trace element solution SL-4 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA | 0.50 | g | ||
|
|
FeSO4 x 7 H2O | 0.20 | g | ||
|
|
ZnSO4 x 7 H2O | 0.10 | g | ||
|
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MnCl2 x 4 H2O | 0.03 | g | ||
|
|
H3BO3 | 0.30 | g | ||
|
|
CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.03 | g | ||
|
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Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA in distilled water and adjust pH to 7.0 using 2 N NaOH; then add other compounds. | |||||
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
|
Thiamine HCl | 50 | mg |
|
|
Riboflavin | 50 | mg |
|
|
Nicotinic acid | 50 | mg |
|
|
Calcium D-(+)-pantothenate | 50 | mg |
|
|
Vitamin B12 | 1 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |