766 .
THERMOCOCCUS CHITINOPHAGUS MEDIUM
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Equipment needed:
Autoclave
Filter
Hungate tubes
Final pH:
6.7
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
(NH4)2SO4 | 0.50 | g | ||
|
|
NaHCO3 | 0.20 | g | ||
|
|
KH2PO4 | 0.50 | g | ||
|
|
Na2WO4 x 2 H2O | 0.15 | mg | ||
|
|
Na2SeO4 | 0.15 | mg | ||
|
|
(NH4)2Ni(SO4)2 | 0.30 | mg | ||
|
|
Chitin stock solution | 4.00 | g | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
Modified Wolin's mineral solution II | 15.00 | ml | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Artifical sea water
(synthetic) |
485.00 | ml | ||
|
|
Distilled water | 500.00 | ml | ||
| 1 Prepare the medium without Na2S x 9 H2O and NaHCO3, boil and cool under N2. Add the NaHCO3 to the cooled medium and adjust the pH to 6.7. Dispense into Hungate tubes or serum bottles under N2 and autoclave. Reduce the medium before use from a sterile stock solution of neutralised Na2S x 9 H2O. | |||||
Chitin stock solution #
1200
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Chitin | 20 | g | ||
|
|
HCl
(37 %) |
200 | ml | ||
|
|
Distilled water | 1000 | ml | ||
| 1 The chitin solution is prepared as follows: mix 20 g chitin (practical grade from crab shells) with 200 ml of 37% HCl (pre-cooled to 4°C) and stir for 1 hr at 4°C. Pour the suspension into 1 litre of distilled water (pre-cooled to 4°C) and filter through filter paper (Schleicher and Schüll Nr. 311853). Wash the residue five times with 500 ml distilled water and resuspend in 1 litre of distilled water. Neutralise the suspension with 10 ml of 5M KOH (final pH 6.5). Filter and wash with 3 litres of distilled water to remove KCl. | |||||
Artifical sea water #
1000
Modified Wolin's mineral solution II #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.500 | g | ||
|
|
MgSO4 x 7 H2O | 3.000 | g | ||
|
|
MnSO4 x H2O | 0.500 | g | ||
|
|
NaCl | 1.000 | g | ||
|
|
FeSO4 x 7 H2O | 0.100 | g | ||
|
|
CoSO4 x 7 H2O | 0.180 | g | ||
|
|
CaCl2 x 2 H2O | 0.100 | g | ||
|
|
ZnSO4 x 7 H2O | 0.180 | g | ||
|
|
CuSO4 x 5 H2O | 0.010 | g | ||
|
|
AlK(SO4)2 x 12 H2O | 0.020 | g | ||
|
|
H3BO3 | 0.010 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.010 | g | ||
|
|
NiCl2 x 6 H2O | 0.025 | g | ||
|
|
Na2SeO3 x 5 H2O | 0.300 | mg | ||
|
|
Distilled water | 1000.000 | 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. | |||||