1245 .
TRICHOCOCCUS SKBG MEDIUM
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Equipment needed:
Anaerobic workbench
Autoclave
Filter
Hungate tubes
Final pH:
7.4
Select a strain (optional):
ml
| 1 Mix components thoroughly and adjust pH to 7.2. Distribute the medium into culture vessels (e.g., 10 ml in Hungate tubes) under a stream of N2-CO2 (4:1, v/v) gas mixture, seal with butyl rubber stoppers and autoclave. Allow to stand overnight. Aseptically and anaerobically add the following solutions (filter-sterilized): | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NaCl | 1.00 | g | ||
|
|
MgCl2 x 6 H2O | 0.20 | g | ||
|
|
NaH2PO4 x 2 H2O | 0.91 | g | ||
|
|
CaCl2 x 2 H2O | 0.10 | g | ||
|
|
NH4Cl | 1.00 | g | ||
|
|
Yeast extract
(BD-Difco) |
0.20 | g | ||
|
|
FeCl2 solution | 1.00 | ml | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Distilled water | 900.00 | ml | ||
|
|
NaHCO3
(8.0%) |
30.00 | ml | ||
|
|
Trace vitamins | 10.00 | ml | ||
|
|
Lactose
(0.25 M) |
50.00 | ml | ||
| 2 Readjust pH to 7.2-7.6, if necessary. | |||||
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 |