611 .
PYROBACULUM (BS) MEDIUM
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Equipment needed:
Autoclave
Gassing station
Serum bottle
Final pH:
7.0 - 7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Synthetic seawater mix | 125.00 | ml | ||
|
|
Modified Wolin's mineral solution | 10.00 | ml | ||
|
|
Fe(NH4)2(SO4)2 x 6 H2O
(0.01% w/v) |
2.00 | ml | ||
|
|
NH4Cl | 0.25 | g | ||
|
|
KH2PO4 | 0.07 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
KNO3 | 1.00 | g | ||
|
|
Yeast extract | 0.50 | g | ||
|
|
NaHCO3 | 2.00 | g | ||
|
|
Distilled water | 875.00 | ml | ||
| 1 Dissolve ingredients (except nitrate, yeast extract and bicarbonate) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Adjust pH to 7.0 and dispense under 100% N2 gas atmosphere into serum vials (e.g., 20 ml medium per 100 ml bottle). Replace gas phase with 80% H2 and 20% CO2 gas mixture and autoclave. Prior to inoculation add nitrate and yeast extract from sterile anoxic stock solutions prepared under 100% N2 gas and bicarbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Adjust pH of complete medium to 7.0 - 7.2. | |||||
Synthetic seawater mix #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaCl | 47.15 | g |
|
|
MgCl2 x 6 H2O | 18.10 | g |
|
|
MgSO4 x 7 H2O | 7.00 | g |
|
|
CaCl2 x 2 H2O | 3.13 | g |
|
|
Na2SO4 | 3.24 | g |
|
|
KCl | 1.20 | g |
|
|
Na2CO3 | 0.10 | g |
|
|
NaBr | 0.10 | g |
|
|
KBr | 80.00 | mg |
|
|
SrCl2 x 6 H2O | 72.00 | mg |
|
|
H3BO3 | 52.00 | mg |
|
|
Na2HPO4 | 8.10 | mg |
|
|
NaF | 2.40 | mg |
|
|
Na-silicate | 0.40 | mg |
|
|
KI | 50.00 | µg |
|
|
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 | ||
|
|
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. | |||||