1508 .
CALDERIHABITANS MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.3 - 7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Sea Salt
(SIGMA) |
21.00 | g | ||
|
|
K2HPO4 | 0.10 | g | ||
|
|
NH4Cl | 0.10 | g | ||
|
|
NaNO3 | 0.05 | g | ||
|
|
Yeast extract | 0.05 | g | ||
|
|
Ferric citrate monohydrate | 0.01 | g | ||
|
|
Trace element solution SL-4 | 1.00 | ml | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Na2CO3 | 1.50 | g | ||
|
|
Na-pyruvate | 1.00 | g | ||
|
|
Na2S2O3 x 5 H2O | 1.00 | g | ||
|
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
Na2S x 9 H2O | 0.25 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except carbonate, pyruvate, thiosulfate, vitamins and sulfide, then sparge medium with 80% N2 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 and autoclave. Add pyruvate, thiosulfate, vitamins 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. Stock solutions of pyruvate, thiosulfate and vitamins should be sterilized by filtration. Prior to use adjust pH of complete medium to 7.3 - 7.5, if necessary. | |||||
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 | ||
|
|
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 | ||
|
|
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 |