1630b .
NITROSOPUMILUS UREIPHILUS MEDIUM
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Equipment needed:
Autoclave
Filter
Final pH:
6.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 26.00 | g | ||
|
|
MgSO4 x 7 H2O | 5.00 | g | ||
|
|
MgCl2 x 6 H2O | 5.00 | g | ||
|
|
CaCl2 x 2 H2O | 1.50 | g | ||
|
|
KBr | 0.10 | g | ||
|
|
KH2PO4
(0.10% w/v) |
2.00 | ml | ||
|
|
NaHCO3 | 0.25 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
NH4Cl | 0.05 | g | ||
|
|
Na-pyruvate | 0.05 | g | ||
|
|
Wolin's vitamin solution | 10.00 | ml | ||
|
|
Catalase
(SIGMA C1345) (0.10% w/v) |
2.50 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except bicarbonate, trace elements, selenite-tungstate, ammonium chloride, pyruvate, vitamins, and catalase) and dispense medium in Erlenmeyer flasks with screw caps (e.g., from SCHOTT), then autoclave. Add bicarbonate, trace elements, selenite-tungstate, ammonium chloride, pyruvate and vitamins from stock solutions sterilized by filtration. Adjust pH of the complete medium to 6.8 using a sterile solution of sodium bicarbonate (10% w/v) or 1 N HCl. | |||||
| 2 After inoculation, add to the medium 5 - 10 units/ml catalase from a freshly prepared 1.00 mg/ml stock solution sterilized by filtration. | |||||
| 3 Note: Use at least 5 - 10% (v/v) inoculum and incubate in the dark without shaking. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium D-(+)-pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |