1204 .
SPHAEROCHAETA MEDIUM
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Selected strain: Sphaerochaeta coccoides DSM 17374
This strain uses the standard medium without modifications.
Growth conditions: 3-7 days
anaerobic
30 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
7.4
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ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.00 | g | ||
|
|
KCl | 0.50 | g | ||
|
|
NH4Cl | 0.30 | g | ||
|
|
KH2PO4 | 0.20 | g | ||
|
|
Na2SO4 | 0.15 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
Yeast extract | 4.00 | g | ||
|
|
Casein peptone | 2.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Na2CO3 | 1.00 | g | ||
|
|
Maltose monohydrate | 2.00 | g | ||
|
|
DL-Dithiothreitol
(DTT) |
0.80 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except carbonate, maltose and DTT, then sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave at 121°C for 20 minutes. Add maltose and DTT from sterile anoxic stock solutions prepared under 100% N2 gas and sterilized by filtration and carbonate from a sterile stock solution prepared under 80% N2 and 20% CO2 gas atmosphere. Check pH of complete medium and adjust to 7.4, if necessary. | |||||