1013 .
HALOBACTERIOVORAX MEDIUM
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Equipment needed:
Autoclave
Final pH:
7.5 - 8.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Artificial sea water | 700 | ml | ||
|
|
Modified Wolin's mineral solution | 10 | ml | ||
|
|
Distilled water | 300 | ml | ||
| 1 Mix ingredients and adjust pH of the final solution to 7.5 - 8.0. Sterilize by autoclaving at 121°C for 20 min. Grow prey bacterium (DSM 15416) on agar plates using medium 514. For 50 ml prey medium suspension use one agar plate incubated for 24 hours and wash off grown cells in 3 - 5 ml medium. The prey medium suspension should contain approx. 107 to 109 cells. Inoculate the complete medium immediately after preparation and incubate the suspension with shaking until a decrease in turbidity is visible. | |||||
Artificial sea water #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 27.70 | g |
|
|
MgSO4 x 7 H2O | 7.00 | g |
|
|
MgCl2 x 6 H2O | 5.50 | g |
|
|
KCl | 0.65 | g |
|
|
NaBr | 0.10 | g |
|
|
H3BO3 | 30.00 | mg |
|
|
SrCl2 x 6 H2O | 15.00 | mg |
|
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Citric acid | 10.00 | mg |
|
|
KI | 0.05 | mg |
|
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CaCl2 x 2 H2O | 2.25 | g |
|
|
Distilled water | 1000.00 | ml |
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
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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 | ||
|
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AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
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H3BO3 | 0.01 | g | ||
|
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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. | |||||
Main sol. 514 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Bacto peptone | 5.00 | g | ||
|
|
Yeast extract
(Bacto ) |
1.00 | g | ||
|
|
Fe(III) citrate | 0.10 | g | ||
|
|
NaCl | 19.45 | g | ||
|
|
MgCl2
(anhydrous) |
5.90 | g | ||
|
|
Na2SO4 | 3.24 | g | ||
|
|
CaCl2 | 1.80 | g | ||
|
|
KCl | 0.55 | g | ||
|
|
NaHCO3 | 0.16 | g | ||
|
|
KBr | 0.08 | g | ||
|
|
SrCl2 | 34.00 | mg | ||
|
|
H3BO3 | 22.00 | mg | ||
|
|
Na-silicate | 4.00 | mg | ||
|
|
NaF | 2.40 | mg | ||
|
|
(NH4)NO3 | 1.60 | mg | ||
|
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Na2HPO4 | 8.00 | mg | ||
|
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Distilled water | 1000.00 | ml | ||
| 1 Final pH should be 7.6 ± 0.2 at 25°C. If using the complete medium from Difco add 37.40 g to 1 litre water. | |||||