143 .
CLOSTRIDIUM SP. LA1 MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final gas composition:
100 % N2
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Final pH:
6.8 - 7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Crotonic acid | 6.00 | g | ||
|
|
NaOH | 0.30 | g | ||
|
|
(NH4)2HPO4 | 0.15 | g | ||
|
|
K2HPO4 | 0.10 | g | ||
|
|
MgCl2 x 6 H2O | 0.03 | g | ||
|
|
CaCl2 x 2 H2O | 0.04 | g | ||
|
|
NH4Cl | 0.05 | g | ||
|
|
MgSO4 x 7 H2O
(0.1% w/v) |
0.60 | ml | ||
|
|
MnSO4 x 2 H2O
(0.1% w/v) |
0.40 | ml | ||
|
|
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
0.40 | ml | ||
|
|
(NH4)6Mo7O24 x 4 H2O | 10.00 | mg | ||
|
|
Biotin
(0.1% w/v) |
0.04 | ml | ||
|
|
p-Aminobenzoic acid
(0.1% w/v) |
0.80 | ml | ||
|
|
Tryptone
(BD BBL) |
1.00 | g | ||
|
|
Yeast extract
(OXOID) |
1.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
K2CO3 | 4.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except potassium carbonate) and 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. An anoxic stock solution of the potassium carbonate is autoclaved separately under 80% N2 and 20% CO2 gas mixture and thereafter added to the sterile medium. Adjust pH of the complete medium to 6.8 - 7.0. Prior to inoculation add 10 - 20 mg/l sodium dithionite from a 5% (w/v) stock solution that has been freshly prepared under 100% N2 gas and sterilized by filtration to reduce the medium. | |||||