1611 .
YCFA MEDIUM (modified)
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Selected strain: Blautia hydrogenotrophica DSM 108220
This strain uses the standard medium without modifications.
Growth conditions: anaerobic
37 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
6.7 - 6.8
Select a strain (optional):
ml
| 1 Prepare the Medium (see Main sol. 1611). Distribute under N2 into Hungate tubes (10 ml): | |||||
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Main sol. 1611 | 10.0 | ml | ||
| 2 Autoclave. Then filter sterilize vitamin solution and aseptically add 100 µl per Hungate tube: | |||||
|
|
Vitamin solution
(filter-sterilized) |
0.1 | ml | ||
Main sol. 1611 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Casitone | 10.00 | g | ||
|
|
Yeast extract | 2.50 | g | ||
|
|
Glucose | 5.00 | g | ||
|
|
MgSO4 x 7 H2O | 45.00 | mg | ||
|
|
CaCl2 x 2 H2O | 90.00 | mg | ||
|
|
K2HPO4 | 0.45 | g | ||
|
|
KH2PO4 | 0.45 | g | ||
|
|
NaCl | 0.90 | g | ||
|
|
Resazurin | 1.00 | mg | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients except NaHCO3, hemin, cysteine in water and boil for 10 min, then add fatty acids: | |||||
|
|
Volatile fatty acids | 2.70 | ml | ||
| 2 Cool down to room temperature while gassing with 100% CO2. Add the NaHCO3, hemin, cysteine. | |||||
|
|
NaHCO3 | 4.00 | g | ||
|
|
L-Cysteine HCl | 1.00 | g | ||
|
|
Hemin | 10.00 | mg | ||
| 3 Adjust pH to 6.7-6.8. | |||||
Volatile fatty acids #
3
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Acetic acid | 1.9 | ml |
|
|
Propionic acid | 0.7 | ml |
|
|
iso-Butyric acid | 90.0 | µl |
|
|
n-Valeric acid | 100.0 | µl |
|
|
iso-Valeric acid | 100.0 | µl |
Vitamin solution #
1000
| Compound | Amount | Unit | |
|---|---|---|---|
|
|
Biotin | 2.0 | mg |
|
|
Folic acid | 2.0 | mg |
|
|
Pyridoxine hydrochloride | 10.0 | mg |
|
|
Thiamine-HCl x 2 H2O | 5.0 | mg |
|
|
Riboflavin | 5.0 | mg |
|
|
Nicotinic acid | 5.0 | mg |
|
|
D-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 |