1604 .
ATHALASSOTOGA MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Ferric citrate monohydrate | 2.40 | g | ||
|
|
(NH4)2SO4 | 0.20 | g | ||
|
|
KH2PO4 | 3.00 | g | ||
|
|
MgSO4 x 7 H2O | 0.50 | g | ||
|
|
CaCl2 x 2 H2O | 0.25 | g | ||
|
|
Yeast extract
(OXOID) |
0.50 | g | ||
|
|
Allen's trace element solution | 10.00 | ml | ||
|
|
FeCl2 x 4 H2O | 0.26 | g | ||
|
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.04 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve ferric citrate in water by heating and adjust to pH 6.0, then add other medium ingredients except ferrous chloride, vitamins and cysteine. Sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic, then dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add ferrous chloride (dissolved in 0.1 N HCl), vitamins (sterilized by filtration) and cysteine from sterile anoxic stock solutions prepared under 100% N2 gas. Prior to inoculation check pH and adjust to 6.0, 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 |