144b .
CALDICELLULOSIRUPTOR OWENSIS MEDIUM
What is this page?
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
7.2 - 7.4
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.90 | g | ||
|
|
NaCl | 0.90 | g | ||
|
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.05 | g | ||
|
|
K2HPO4 | 0.40 | g | ||
|
|
Trace element solution | 10.00 | ml | ||
|
|
Yeast extract | 3.00 | g | ||
|
|
Trypticase peptone
(BD BBL) |
10.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Na2CO3 | 2.00 | g | ||
|
|
Wolin's vitamin solution | 5.00 | ml | ||
|
|
D-Glucose | 5.00 | g | ||
|
|
Na2S x 9 H2O | 1.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except carbonate, vitamins, glucose and sulfide), sparge medium with 80% N2 and 20% CO2 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. Add glucose, vitamins (sterilized by filtration) 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. Adjust pH of complete medium to 7.2 - 7.4. | |||||
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2-EDTA x 2 H2O | 0.50 | g | ||
|
|
CoCl2 x 6 H2O | 0.15 | g | ||
|
|
MnCl2 x 4 H2O | 0.10 | g | ||
|
|
FeSO4 x 7 H2O | 0.10 | g | ||
|
|
ZnCl2 | 0.10 | g | ||
|
|
AlCl3 x 6 H2O | 40.00 | mg | ||
|
|
Na2WO4 x 2 H2O | 40.00 | mg | ||
|
|
Na2SeO3 x 5 H2O | 30.00 | mg | ||
|
|
NiCl2 x 6 H2O | 20.00 | mg | ||
|
|
CuCl2 x 2 H2O | 20.00 | mg | ||
|
|
H3BO3 | 10.00 | mg | ||
|
|
Na2MoO4 x 2 H2O | 10.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 First dissolve EDTA in distilled water and adjust pH to 7 using 2 N NaOH, then dissolve remaining compounds. | |||||
Wolin's vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine HCl | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
Calcium D-(+)-pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |