920 .
HYDROGENISPORA MEDIUM
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Equipment needed:
Final pH:
6.5
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract
(BD-Difco) |
0.20 | g | ||
|
|
KH2PO4 | 0.14 | g | ||
|
|
MgCl2 x 6 H2O | 0.20 | g | ||
|
|
CaCl2 x 2 H2O | 0.15 | g | ||
|
|
NH4Cl | 0.54 | g | ||
|
|
Trace vitamins solution | 1.00 | ml | ||
|
|
Trace element solution | 1.00 | ml | ||
|
|
Se/W solution | 1.00 | ml | ||
|
|
NaHCO3 | 2.50 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Mix components, except NaHCO3, and adjust pH to 6.5. Bring to a boil for 5-10 sec, cool down under a stream of a N2-CO2 (80:20 v/v) gas mixture and add NaHCO3 to the solution. Distribute the medium into culture vessels (e.g., 20 ml medium in 60 ml serum bottle) under the same gas mixture and seal with butyl rubber stoppers. After autoclaving, stand the medium overnight. Then, add per liter the following solutions: | |||||
|
|
Glucose
(1.0 M) |
10.00 | ml | ||
|
|
L-Cysteine HCl x H2O
(5%) |
6.00 | ml | ||
|
|
Na2S x 9 H2O
(5%) |
6.00 | ml | ||
Trace vitamins solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 4.9 | mg |
|
|
Folic acid | 8.8 | mg |
|
|
Pyridoxine hydrochloride | 4.1 | mg |
|
|
Thiamine HCl | 6.7 | mg |
|
|
Riboflavin | 7.5 | mg |
|
|
Nicotinic acid | 2.5 | mg |
|
|
DL-Calcium pantothenate | 9.5 | mg |
|
|
Vitamin B12 | 27.1 | mg |
|
|
p-Aminobenzoic acid | 2.7 | mg |
|
|
Lipoic acid | 4.1 | mg |
|
|
Distilled water | 1000.0 | ml |
Trace element solution #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Nitrilotriacetic acid | 12.800 | g | ||
|
|
FeCl3 x 6 H2O | 1.350 | g | ||
|
|
MnCl2 x 4 H2O | 0.100 | g | ||
|
|
CoCl2 x 6 H2O | 0.024 | g | ||
|
|
CaCl2 x 2 H2O | 0.100 | g | ||
|
|
ZnCl2 | 0.100 | g | ||
|
|
CuCl2 x 2 H2O | 0.025 | g | ||
|
|
H3BO3 | 0.010 | g | ||
|
|
Na2MoO4 x 2 H2O | 0.024 | g | ||
|
|
NiCl2 x 6 H2O | 0.120 | g | ||
|
|
Distilled water | 1000.000 | ml | ||
| 1 Dissolve nitrilotriacetic acid first in 200 ml of distilled water and adjust pH to 6.5 with KOH. Then add the remaining salts and bring volume to 1.0 L. Readjust pH to 7.0 with KOH. | |||||
Se/W solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Na2SeO3 x 5 H2O | 4 | mg |
|
|
Na2WO4 x 2 H2O | 4 | mg |
|
|
Distilled water | 1000 | ml |