1690 .
NITROSPIRA INOPINATA MEDIUM
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Equipment needed:
Autoclave
Final pH:
7.6 - 8.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 1.00 | g | ||
|
|
KCl | 1.50 | g | ||
|
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MgSO4 x 7 H2O | 1.00 | g | ||
|
|
NaCl | 11.68 | g | ||
|
|
Distilled water
(MilliQ) |
1000.00 | ml | ||
|
|
CaCO3 | 4.00 | g | ||
| 1 Dilute stock solution with MilliQ water (1+19 v/v), add 4 g CaCO3 per litre and autoclave. After autoclaving add sterile solutions: | |||||
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Trace element solution SL-10 | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
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Borate-copper solution | 1.00 | ml | ||
|
|
Ammonium chloride solution | 10.00 | ml | ||
| 2 Complete medium can be stored at 4°C. | |||||
| 3 Check pH 7.6-8.0 | |||||
Borate-copper solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
H3BO3 | 54 | mg | ||
|
|
CuCl2 x 2 H2O | 18 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Incubate at 42°C without shaking. Activity and growth can be followed by measuring the depletion of ammonium or the production of nitrite and/or nitrate. Higher ammonium concentrations are inhibitive. For that reason our recommendation is follow nitrite content. As soon as nitrite has been produced and is depleting again (to form nitrate) another dose of ammonium can be added. Incubation time may be up to several weeks. We checked the nitrite content (absence/presence) every 5-7 days and added ammonium chloride about every 2-3 weeks. Don't expose cultures to intense light. | |||||
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 | ||
|
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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 |
Ammonium chloride solution #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Ammonium chloride | 0.8 | g |
|
|
Distilled water | 100.0 | ml |