432 .
HALOBACTEROIDES ACETOETHYLICUS MEDIUM
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Selected strain: Halanaerobium acetethylicum DSM 3532
This strain uses the standard medium without modifications.
Growth conditions: 30 °C
Equipment needed:
Autoclave
Filter
Gassing station
Hungate tubes
Final pH:
n.d.
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Medium 144 | 1000 | ml | ||
|
|
NaCl | 100 | g/l | ||
| 1 Use medium no. 144 supplemented with 100 g/l of NaCl. | |||||
| 2 Sterilize glucose, yeast extract and sodium sulfide separately (autoclave under nitrogen atmosphere). | |||||
Main sol. 144 #
1017
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
NH4Cl | 0.90 | g | ||
|
|
NaCl | 0.90 | g | ||
|
|
MgCl2 x 6 H2O | 0.40 | g | ||
|
|
KH2PO4 | 0.75 | g | ||
|
|
K2HPO4 | 1.50 | g | ||
|
|
Trace element solution | 9.00 | ml | ||
|
|
FeSO4 x 7 H2O
(0.1% w/v in 0.1 N H2SO4) |
3.00 | ml | ||
|
|
Yeast extract | 3.00 | g | ||
|
|
Trypticase peptone
(BD BBL) |
10.00 | g | ||
|
|
Sodium resazurin
(0.1% w/v) |
0.50 | ml | ||
|
|
Wolin's vitamin solution | 5.00 | ml | ||
|
|
D-Glucose | 5.00 | g | ||
|
|
Na2S x 9 H2O | 1.00 | g | ||
|
|
Distilled water | 1000.00 | ml | ||
| 1 Dissolve ingredients (except vitamins, glucose and sulfide), sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Dispense medium under same gas atmosphere into anoxic Hungate-type tubes or serum vials and autoclave. Add glucose, vitamins (sterilized by filtration) and sulfide from sterile anoxic stock solutions prepared under 100% N2 gas. Adjust pH of complete medium to 7.2 - 7.4. | |||||
Wolin's vitamin solution #
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 D-(+)-pantothenate | 5.0 | mg |
|
|
Vitamin B12 | 0.1 | mg |
|
|
p-Aminobenzoic acid | 5.0 | mg |
|
|
(DL)-alpha-Lipoic acid | 5.0 | mg |
|
|
Distilled water | 1000.0 | ml |