1479 .
HALODESULFURIARCHAEUM MEDIUM
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Equipment needed:
Autoclave
Filter
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 240.0 | g | ||
|
|
K2HPO4 | 2.5 | g | ||
|
|
(NH4)2SO4 | 0.5 | g | ||
|
|
HEPES | 10.0 | g | ||
|
|
Trace element solution | 1.0 | ml | ||
|
|
Sulfur powder | 2.0 | g | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 Prepare the liquid medium. Dispense the medium into Hungate tubes or serum bottles under gassing with N2. Steam medium for 3h on each of 3 successive days. Before inoculation add from sterile stock solutions: | |||||
|
|
MgCl2 x 6 H2O | 3.0 | mM | ||
|
|
NaHCO3 | 5.0 | mM | ||
|
|
Sodium formate | 50.0 | mM | ||
|
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Yeast extract | 0.2 | g/l | ||
|
|
Vitamin mix (pfennig & Lippert,1966) | 1.0 | ml | ||
|
|
Disodium sulfide
(1M filter sterilized stock) |
1.0 | mM | ||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
EDTA | 5.00 | g | ||
|
|
FeSO4 x 7 H2O | 2.00 | g | ||
|
|
ZnSO4 | 0.10 | g | ||
|
|
MnCl2 | 0.03 | g | ||
|
|
H3BO3 | 0.30 | g | ||
|
|
CoCl2 x 6 H2O | 0.20 | g | ||
|
|
CuCl2 x 5 H2O | 0.01 | g | ||
|
|
NiCl2 x 6 H2O | 0.02 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.02 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Adjust pH to 3.0-4.0 with HCl. | |||||
| 2 Autoclave at 121 °C for 15 min. | |||||
| 3 Note: autoclave in closed bottles | |||||
Vitamin mix (pfennig & Lippert,1966) #
400
| 1 1: acidic solution (HCl, pH3; 100ml) | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Thiamin | 5.0 | mg | ||
|
|
Calcium pantothenate | 5.0 | mg | ||
| 2 2: basic solution (100ml 0.1N NaOH) | |||||
|
|
Biotin | 2.0 | mg | ||
|
|
PABA
(p-aminobenzoic acid) |
5.0 | mg | ||
|
|
Nicotinic acid | 5.0 | mg | ||
|
|
Pyridoxine | 10.0 | mg | ||
| 3 3: Neutral solution in 100 ml demi water | |||||
|
|
Folic acid | 2.0 | mg | ||
|
|
Riboflavin | 5.0 | mg | ||
| 4 4: neutral solution in 100 ml demi water | |||||
|
|
B12 | 0.5 | mg | ||
| 5 Sterilize by filtration. | |||||
| 6 Mix 1:1:1:1 just after addition or add separatlely, each 1ml/L. | |||||