289 .
RUMINICLOSTRIDIUM PAPYROSOLVENS MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
7.2
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Sea water
(filtered) |
200.00 | ml | ||
|
|
Mineral salt solution | 150.00 | ml | ||
|
|
NH4Cl | 1.00 | g | ||
|
|
Yeast extract | 0.60 | g | ||
|
|
K2HPO4 | 1.65 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Cellobiose | 5.00 | g | ||
|
|
Cellulose
, Lens tissue or MN 301
optional |
5.00 | g | ||
|
|
Distilled water | 650.00 | ml | ||
| 1 Dissolve ingredients (except cysteine and cellobiose) and sparge medium with 100% N2 gas for 30 - 45 m in to make it anoxic. Add cysteine, adjust pH to 7.2 and dispense under 100% N2 gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. After sterilization add cellobiose from an anoxic stock solution sterilized by filtration. | |||||
| 2 Note: Some strains can be adapted to cellulose as substrate using 5.0 g/l cellulose (Lens tissue, OXOID; or cellulose powder MN 301, MACHEREY-NAGEL). If necessary adjust pH of completed medium to 7.2. | |||||
Mineral salt solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
(NH4)2SO4 | 6.0 | g |
|
|
NaCl | 6.0 | g |
|
|
MgSO4 x 7 H2O | 1.2 | g |
|
|
CaCl2 x 2 H2O | 0.8 | g |
|
|
Distilled water | 1000.0 | ml |