727 .
CLOSTRIDIUM ULTUNENSE MEDIUM
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Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
KH2PO4 | 0.40 | g | ||
|
|
Na2HPO4 | 0.43 | g | ||
|
|
NH4Cl | 0.30 | g | ||
|
|
NaCl | 0.30 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Selenite-tungstate solution | 1.00 | ml | ||
|
|
Yeast extract | 10.00 | g | ||
|
|
Peptone | 10.00 | g | ||
|
|
Lab Lemco powder
(Oxoid) |
5.00 | g | ||
|
|
Starch
(soluble) |
0.40 | g | ||
|
|
Na-acetate | 0.40 | g | ||
|
|
Resazurin | 0.50 | mg | ||
|
|
Na2CO3 | 1.50 | g | ||
|
|
Wolin's vitamin solution (10x) | 1.00 | ml | ||
|
|
Na2S x 9 H2O | 0.30 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except carbonate, vitamins and sulfide, then sparge medium with 80% N2 and 20% CO2 gas mixture for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Prior to inoculation add vitamins and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas and carbonate from a sterile anoxic stock solution prepared under 80% N2 and 20% CO2 gas mixture. Stock solutions of vitamins should be sterilized by filtration. Adjust pH of complete medium to 7.0, if necessay. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA | 0.5 | g | ||
|
|
FeCl2 x 4 H2O | 2.0 | g | ||
|
|
ZnCl2 | 70.0 | mg | ||
|
|
MnCl2 x 4 H2O | 100.0 | mg | ||
|
|
H3BO3 | 6.0 | mg | ||
|
|
CoCl2 x 6 H2O | 190.0 | mg | ||
|
|
CuCl2 x 2 H2O | 2.0 | mg | ||
|
|
AlCl3 | 10.0 | mg | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 1000.0 | ml | ||
| 1 First dissolve EDTA in distilled water, adjust pH to 7 using 2 N NaOH and add ferrous chloride. After ferrous chloride has dissolved add remaining compounds. | |||||
Selenite-tungstate solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 0.5 | g |
|
|
Na2SeO3 x 5 H2O | 3.0 | mg |
|
|
Na2WO4 x 2 H2O | 4.0 | mg |
|
|
Distilled water | 1000.0 | ml |
Wolin's vitamin solution (10x) #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 20 | mg |
|
|
Folic acid | 20 | mg |
|
|
Pyridoxine hydrochloride | 100 | mg |
|
|
Thiamine HCl | 50 | mg |
|
|
Riboflavin | 50 | mg |
|
|
Nicotinic acid | 50 | mg |
|
|
Calcium D-(+)-pantothenate | 50 | mg |
|
|
Vitamin B12 | 1 | mg |
|
|
p-Aminobenzoic acid | 50 | mg |
|
|
(DL)-alpha-Lipoic acid | 50 | mg |
|
|
Distilled water | 1000 | ml |