1360 .
HIPPEA JASONIAE MEDIUM
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Equipment needed:
Filter
Gassing station
Serum bottle
Water bath
Final pH:
4.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 26.33 | g | ||
|
|
MgCl2 x 6 H2O | 1.50 | g | ||
|
|
KCl | 0.45 | g | ||
|
|
(NH4)2SO4 | 1.30 | g | ||
|
|
KH2PO4 | 0.28 | g | ||
|
|
MgSO4 x 7 H2O | 0.25 | g | ||
|
|
CaCl2 x 2 H2O | 0.07 | g | ||
|
|
FeCl3 x 6 H2O | 0.02 | g | ||
|
|
Allen's trace element solution | 10.00 | ml | ||
|
|
Na-acetate | 1.25 | g | ||
|
|
Trisodium citrate x 2 H2O | 2.94 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Sulfur , powdered | 10.00 | g | ||
|
|
Yeast extract
(DIFCO) |
0.50 | g | ||
|
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except sulfur, vitamins, yeast extract and sulfide), adjust pH to 3.5 with 10 N H2SO4 and 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 serum vials (e.g., 20 ml medium in 100 ml bottles) that contain already the appropriate amount of sulfur. Sterilize medium by heating cultivation vessels in a water bath to 90 - 100°C for 1 - 2 hours on each of 3 successive days. Add vitamins (sterilized by filtration) and yeast extract from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to inoculation, add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas and adjust pH of complete medium to 4.5, if necessary. | |||||
Allen's trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
MnCl2 x 4 H2O | 180 | mg | ||
|
|
Na2B4O7 x 10 H2O | 450 | mg | ||
|
|
ZnSO4 x 7 H2O | 22 | mg | ||
|
|
CuCl2 x 2 H2O | 5 | mg | ||
|
|
Na2MoO4 x 2 H2O | 3 | mg | ||
|
|
VOSO4 x 2 H2O | 3 | mg | ||
|
|
CoSO4 x 7 H2O | 1 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Adjust pH of final solution to 2 with 1 N HCl. | |||||
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 |