945 .
MARINITOGA PIEZOPHILA MEDIUM
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Equipment needed:
Gassing station
Hungate tubes
Water bath
Final gas composition:
100 % N2
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Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 1.00 | g | ||
|
|
MgCl2 x 6 H2O | 0.20 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
KCl | 0.10 | g | ||
|
|
NaCl | 30.00 | g | ||
|
|
Na-acetate | 0.83 | g | ||
|
|
MES
(SIGMA) |
1.95 | g | ||
|
|
Yeast extract | 5.00 | g | ||
|
|
Trypticase peptone
(BD BBL) |
5.00 | g | ||
|
|
K2HPO4 | 0.30 | g | ||
|
|
KH2PO4 | 0.30 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Sulfur
(powdered) |
10.00 | g | ||
|
|
L-Cysteine HCl x H2O | 0.30 | g | ||
|
|
Na2S x 9 H2O | 0.30 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except sulfur, cysteine and sulfide. Sparge medium with 100% N2 gas for at least 30 - 45 min to make it anoxic and adjust pH to 6.0 with NaOH. Distribute the medium under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials which contain already the appropriate amount of sulfur. For sterilization heat the cultivation vessels containing medium plus sulfur in boiling water for 2 - 3 hours on each of 3 successive days! Prior to inoculation add cysteine and sulfide and from sterile anoxic stock solutions prepared under 100% N2 gas. | |||||
| 2 After inoculation pressurize tubes to 2 bar overpressure with sterile 100% N2 gas. | |||||