1083 .
DESULFURELLA TR1 MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.0
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Na2HPO4 x 2 H2O | 0.53 | g | ||
|
|
KH2PO4 | 0.41 | g | ||
|
|
NH4Cl | 0.30 | g | ||
|
|
NaCl | 0.30 | g | ||
|
|
MgCl2 x 6 H2O | 0.10 | g | ||
|
|
CaCl2 x 2 H2O | 0.11 | g | ||
|
|
Acid trace element solution | 1.00 | ml | ||
|
|
Alkaline trace element solution | 1.00 | ml | ||
|
|
Sodium acetate x 3 H2O | 0.68 | g | ||
|
|
Yeast extract | 0.10 | g | ||
|
|
Sulfur
(powder) |
0.80 | g | ||
|
|
Resazurin | 1.00 | m | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components, except sulfur, and autoclave under a N2-CO2 (4:1, v/v) gas mixture. After cooling, add the following solutions (filter-sterilized): | |||||
|
|
Vitamin solution | 0.20 | ml | ||
|
|
NaHCO3
(8%) |
12.50 | ml | ||
| 2 Steam sulfur for 3 hr on each of 3 successive days. Aseptically and anaerobically distribute the medium and sulfur into culture vessels (e.g., 50 ml of the medium and 0.04 g of sulfur in 120 ml serum bottle) under a N2-CO2 (4:1, v/v) gas mixture, and seal with butyl rubber stoppers. Prior to use, anaerobically add per liter the following solution from an anaerobic stock (autoclaved and stored under a N2 atmosphere): | |||||
|
|
Na2S x 9 H2O
(5%) |
6.00 | ml | ||
| 3 Adjust pH to 6.0, if necessary. | |||||
Acid trace element solution #
1010
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(6 N) |
10.00 | ml |
|
|
H3BO3 | 61.80 | mg |
|
|
MnCl2 x 4 H2O | 61.25 | mg |
|
|
FeCl2 x 4 H2O | 943.50 | mg |
|
|
CoCl2 x 6 H2O | 64.50 | mg |
|
|
NiCl2 x 6 H2O | 12.86 | mg |
|
|
ZnCl2 | 67.70 | mg |
|
|
Distilled water | 1000.00 | ml |
Alkaline trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
NaOH | 400.0 | mg |
|
|
Na2SeO3 x 5 H2O | 17.3 | mg |
|
|
Na2WO4 x 2 H2O | 29.4 | mg |
|
|
Na2MoO4 x 2 H2O | 20.5 | mg |
|
|
Distilled water | 1000.0 | ml |
Vitamin solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Biotin | 20 | mg | ||
|
|
Nicotinic acid | 200 | mg | ||
|
|
p-Aminobenzoic acid | 100 | mg | ||
|
|
Thiamine HCl | 200 | mg | ||
|
|
DL-Calcium pantothenate | 100 | mg | ||
|
|
Pyridoxine hydrochloride | 500 | mg | ||
|
|
Vitamin B12 | 100 | mg | ||
|
|
Riboflavin | 100 | mg | ||
|
|
Distilled water | 1000 | ml | ||
| 1 Filter-sterilize and store under a N2 atmosphere. | |||||