795 .
STETTERIA MEDIUM
What is this page?
Equipment needed:
Gassing station
Serum bottle
Water bath
Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Synthetic seawater (2 x conc.) | 500.00 | ml | ||
|
|
NaHCO3 | 0.16 | g | ||
|
|
KH2PO4 | 0.50 | g | ||
|
|
Modified Wolin's mineral solution | 15.00 | ml | ||
|
|
NiCl2 x 6 H2O
(0.1% w/v) |
3.00 | ml | ||
|
|
Peptone
(BD Bacto) |
2.00 | g | ||
|
|
Yeast extract
(OXOID) |
1.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Sulfur
, powdered
(powdered) |
10.00 | g | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
Distilled water | 500.00 | ml | ||
| 1 Dissolve ingredients except sulfur and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into serum vials (e.g., 20 ml medium per 100 ml bottle) that contain already the appropriate amount of sulfur. After closing pressurize bottles to 2 bar overpressure using 80% H2 and 20% CO2 gas mixture, than heat at 100°C for 1.5 hours on each of 3 successive days. Before use add sulfide from a sterile anoxic stock solution prepared under 100% N2 gas atmosphere. Adjust pH of complete medium to 6.0, if necessary. Incubate without shaking. | |||||
Modified Wolin's mineral solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 1.50 | g | ||
|
|
MgSO4 x 7 H2O | 3.00 | g | ||
|
|
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 | ||
|
|
AlK(SO4)2 x 12 H2O | 0.02 | g | ||
|
|
H3BO3 | 0.01 | g | ||
|
|
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. | |||||
Synthetic seawater (2 x conc.) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 55.40 | g |
|
|
SrCl2 x 6 H2O | 0.03 | g |
|
|
MgSO4 x 7 H2O | 14.00 | g |
|
|
KI | 0.10 | mg |
|
|
MgCl2 x 6 H2O | 11.00 | g |
|
|
Na3-citrate | 20.00 | mg |
|
|
KCl | 1.30 | g |
|
|
CaCl2 x 2 H2O | 1.50 | g |
|
|
NaBr | 0.20 | g |
|
|
H3BO3 | 0.06 | g |
|
|
Distilled water | 1000.00 | ml |