1009 .
CLOSTRIDIUM SWELLFUNIANUM MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
7.3
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.33 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
KH2PO4 | 0.33 | g | ||
|
|
Yeast extract
(BD-Difco) |
0.50 | g | ||
|
|
Tryptone
(BD-Difco) |
3.00 | g | ||
|
|
Trace element solution SL-10 | 1.00 | ml | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
CaCl2 x 2 H2O
(1%) |
33.00 | ml | ||
|
|
MgCl2 x 6 H2O
(2%) |
16.50 | ml | ||
|
|
L-Cysteine HCl x H2O | 0.50 | g | ||
|
|
Resazurin
(1%) |
1.00 | ml | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Prepare the medium anaerobically under a N2-CO2 (4:1, v/v) gas mixture. Distribute the medium into culture vessels (e.g., 5.0 ml in Hungate tubes) under the same gas mixture, seal with butyl rubber stoppers and autoclave. After autoclaving, add per 5.0 ml the following solutions from anaerobic stocks (autoclaved or *filter-sterilized and stored under a N2 atmosphere): | |||||
|
|
Na2S x 9 H2O
(w/v, 3%) |
0.05 | ml | ||
|
|
NaHCO3
(w/v, *, 10%) |
0.05 | ml | ||
|
|
Glucose
(w/v, 20%) |
0.10 | ml | ||
| 2 Adjust pH to 7.3. | |||||
Trace element solution SL-10 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
HCl
(25%, 7.7 M) |
10.0 | ml | ||
|
|
FeCl2 x 4 H2O | 1.5 | 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 | ||
|
|
NiCl2 x 6 H2O | 24.0 | mg | ||
|
|
Na2MoO4 x 2 H2O | 36.0 | mg | ||
|
|
Distilled water | 990.0 | ml | ||
| 1 Dissolve FeCl2 x 4H2O first in HCl. Then dilute in distilled water and add the remaining salts. | |||||
Trace vitamins #
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 pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |