1464 .
CHITINIVIBRIO MEDIUM
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* Medium for Chitinivibrio alkaliphilus DSM 24538
Use chitin C7170 by Sigma, others may not work as well
Growth conditions: anaerobic
30 °C
Equipment needed:
Autoclave
Final pH:
10.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaHCO3 | 15.0 | g/l | ||
|
|
Na2CO3 | 95.0 | g/l | ||
|
|
NaCl | 6.0 | g/l | ||
|
|
K2HPO4 | 1.0 | g/l | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Adjust pH to 10.0 with NaOH or HCl. Make anoxic under nitrogen stream and autoclave at 121°C. | |||||
| 2 After autoclaving add sterile, anoxic solutions of: | |||||
|
|
Chitin
, adjust volume in range 1-2 if required
(amorphous or Sigma C7170, 5% solution) |
1.5 | g | ||
|
|
MgSO4 | 1.0 | mM | ||
|
|
Yeast extract | 20.0 | mg | ||
|
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NH4Cl | 4.0 | mM | ||
|
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Na2S | 1.0 | mM | ||
|
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Vitamine mixture (Pfennig & Lippert, 1965) | 1.0 | ml | ||
|
|
Acidic trace metals (Pfennig & Lippert, 1965) | 1.0 | ml | ||
Vitamine mixture (Pfennig & Lippert, 1965) #
150
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Basic solution (100 ml 0.1 N NaOH) | 50 | ml | ||
|
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Neutral solution (100 ml Aqua dem.) | 50 | ml | ||
|
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Acidic solution (HCl, pH 3; 100 ml) | 50 | ml | ||
| 1 Sterilize by filtration and keep at 4°C. Before addition to the medium mix 1:1:1 (vol:vol:vol). | |||||
Basic solution (100 ml 0.1 N NaOH) #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2 | mg |
|
|
PABA | 5 | mg |
|
|
Nicotinic acid | 5 | mg |
|
|
Pyridoxine | 10 | mg |
|
|
Distilled water | 100 | ml |
Neutral solution (100 ml Aqua dem.) #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Folic acid | 2 | mg |
|
|
Riboflavin | 5 | mg |
|
|
Distilled water | 100 | ml |
Acidic trace metals (Pfennig & Lippert, 1965) #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5 | g/l | ||
|
|
FeSO4 x 7 H2O | 2 | g/l | ||
|
|
ZnSO4 x 7 H2O | 100 | mg/l | ||
|
|
MnCl2 | 30 | mg/l | ||
|
|
H3BO3 | 300 | mg/l | ||
|
|
CoCl2 x 6 H2O | 200 | mg/l | ||
|
|
CuCl2 | 10 | mg/l | ||
|
|
NiCl2 x 2 H2O | 20 | mg/l | ||
|
|
Na2MoO4 x 2 H2O | 20 | mg/l | ||
|
|
Distilled water | 1000 | ml | ||
| 1 adjust pH to 3-4 with HCl; autoclave in closed bottles, 120°C, 20 min. | |||||
Acidic solution (HCl, pH 3; 100 ml) #
100
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamin | 5.0 | mg |
|
|
Calcium pantothenate | 5.0 | mg |
|
|
B12 | 0.5 | mg |
|
|
Distilled water | 100.0 | ml |