1290 .
ALKALIPHILUS MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
8.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Solution A | 950 | ml | ||
|
|
Solution B | 25 | ml | ||
|
|
Solution C | 1 | ml | ||
|
|
Solution D | 25 | ml | ||
| 1 Dissolve ingredients of solution A, adjust pH to 8.0 with 1 N NaOH and boil medium for 1 min, then cool to room temperature under 100% N2 gas atmosphere. Dispense solution A under same gas atmosphere in anoxic culture vessels (e.g., Hungate-type tubes) and autoclave. Prior to inoculation complete the medium by adding appropriate amounts of solutions B, C and D prepared under 100% N2 gas and sterilized by filtration. Adjust pH of the completed medium to 8.0 with a sterile anoxic solution of 0.1 N HCl or 0.1 N NaOH, if necessary. | |||||
Solution A #
950
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 75.0 | g |
|
|
MgSO4 x 7 H2O | 8.8 | g |
|
|
KCl | 1.0 | g |
|
|
Yeast extract
(OXOID) |
10.0 | g |
|
|
Trypticase peptone
(BD BBL) |
10.0 | g |
|
|
Modified Wolin's mineral solution | 10.0 | ml |
|
|
Distilled water | 940.0 | ml |
Solution D #
25
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na-thioglycolate | 0.5 | g |
|
|
Na-ascorbate | 0.5 | g |
|
|
Distilled water | 25.0 | ml |
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
MnSO4 x H2O | 0.50 | g | ||
|
|
NaCl | 1.00 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
CoSO4 x 7 H2O | 0.18 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
ZnSO4 x 7 H2O | 0.18 | g | ||
|
|
CuSO4 x 5 H2O | 0.01 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.03 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.30 | mg | ||
|
|
Na2WO4 x 2 H2O | 0.40 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve nitrilotriacetic acid and adjust pH to 6.5 with KOH, then add minerals. Adjust final to pH 7.0 with KOH. | |||||
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 |