1309 .
3.8M CHARACTERIZATION MEDIUM NO. 1
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Equipment needed:
Autoclave
Filter
Final pH:
7.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Salt water
(see below, 30%) |
830.000 | ml | ||
|
|
Peptone
(Oxoid) |
0.025 | g | ||
|
|
Yeast extract | 5.000 | mg | ||
| 1 Mix components and autoclave. After cooling, add the following solutions (autoclaved or *filter-sterilized): | |||||
|
|
NH4Cl
(1M) |
5.000 | ml | ||
|
|
K2HPO4
(0.5 M) |
2.000 | ml | ||
|
|
FeSO4 x 7 H2O
(*, 0.15%) |
5.000 | ml | ||
|
|
Trace element solution | 5.000 | ml | ||
|
|
Vitamin solution
(*) |
3.000 | ml | ||
|
|
Sodium pyruvate
(1.0 M) |
50.000 | ml | ||
|
|
Sodium perchlorate
(1.0 M) |
50.000 | ml | ||
|
|
Sodium Thiosulfate
(1.0 M) |
50.000 | ml | ||
| 2 30% Salt water: | |||||
|
|
NaCl | 240.000 | g | ||
|
|
MgCl2 x 6 H2O | 24.900 | g | ||
|
|
MgSO4 x 7 H2O | 29.000 | g | ||
|
|
KCl | 7.000 | g | ||
|
|
CaCl2
(1M) |
5.000 | ml | ||
| 3 Aseptically distribute the medium into culture vessels (e.g., 10 ml medium in Balch tubes), seal with butyl rubber stoppers and replace the gas phase with N2. | |||||
| 4 Add components to distilled water and bring volume to 1.0 L. Adjust pH to 7.5. | |||||
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Thiamine HCl | 33 | mg |
|
|
Riboflavin | 10 | mg |
|
|
Nicotinic acid | 33 | mg |
|
|
DL-Calcium pantothenate | 17 | mg |
|
|
Vitamin B12 | 17 | mg |
|
|
p-Aminobenzoic acid | 13 | mg |
|
|
Distilled water | 1000 | 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 |