187 .
DESULFUROCOCCUS AMYLOLYTICUS MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Final pH:
6.3
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.33 | g | ||
|
|
KH2PO4 | 0.33 | g | ||
|
|
KCl | 0.33 | g | ||
|
|
CaCl2 x 2 H2O | 0.44 | g | ||
|
|
MgCl2 x 6 H2O | 0.70 | g | ||
|
|
NaCl | 0.50 | g | ||
|
|
NaHCO3 | 0.80 | g | ||
|
|
FeCl2 solution | 1.00 | ml | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
Yeast extract
(BD-Difco) |
0.20 | g | ||
|
|
Bacto peptone
(BD-Difco) |
5.00 | g | ||
|
|
Sulfur
(powder) |
10.00 | g | ||
|
|
Na2S x 9 H2O | 0.50 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 990.00 | ml | ||
| 1 Mix ingredients, except yeast extract, peptone, vitamins, sulfur and Na2S x 9H2O, and autoclave under a N2-CO2 (80:20) atmosphere. Separately autoclave yeast extract and peptone as 10% solutions and Na2S x 9H2O as a 5% solution under a N2 atmosphere. Steam sulfur for 3 hr on each of 3 successive days. Filter-sterilize NaHCO3 as an 8% solution and vitamin solution while gassing with N2. Aseptically and anaerobically add yeast extract, peptone, NaHCO3 and vitamin solutions, and sulfur to the medium under a N2-CO2 (80:20) atmosphere. Prior to inoculation, add the sterile Na2S x 9H2O solution, and adjust pH to 6.2-6.4 with sterile 1 N H2SO4. | |||||
FeCl2 solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
HCl
(7.7 M, 25%) |
10.0 | ml |
|
|
FeCl2 x 4 H2O | 1.5 | g |
|
|
Distilled water | 990.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
ZnCl2 | 70 | mg |
|
|
MnCl2 x 4 H2O | 100 | mg |
|
|
H3BO3 | 6 | mg |
|
|
CoCl2 x 6 H2O | 190 | mg |
|
|
CuCl2 x 2 H2O | 2 | mg |
|
|
NiCl2 x 6 H2O | 24 | mg |
|
|
Na2MoO4 x 2 H2O | 36 | mg |
|
|
Distilled water | 1000 | ml |
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 |