848 .
RAE MEDIUM
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Selected strain: Glutamicibacter mysorens DSM 12798
This strain uses the standard medium without modifications.
Equipment needed:
Autoclave
Filter
Final pH:
3.8
Select a strain (optional):
ml
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
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Glucose | 40.00 | g | ||
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Peptone | 10.00 | g | ||
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Yeast extract | 10.00 | g | ||
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Citric acid x H2O | 1.50 | g | ||
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Na2HPO4 x 2 H2O | 3.38 | g | ||
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Glacial acetic acid | 10.00 | ml | ||
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Absolute ethanol | 10.00 | ml | ||
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Distilled water | 980.00 | ml | ||
| 1 Final pH ca. 3.8 (pH is not adjusted) | |||||
| 2 Both liquid and solid media are prepared by autoclaving the medium without the addition of glacial acetic acid and absolute ethanol. Glacial acetic acid and absolute ethanol are sterilised either by filtration (use Teflon filters) or they may be autoclaved in completely closed screw cap glass bottles sealed with a Teflon coated septum. The preparation of solid media employs a double layer system similar to that described for Acetobacter europaeus medium (see medium 631). Substitute the components listed above and prepare a lower, 0.5% agar layer, which is then overlaid with a 1.0% agar layer as described in medium 631. Incubate in closed containers to keep the humidity high (> 90%). | |||||
Main sol. 631 #
1000
| Compound | Amount | Unit | |||
|---|---|---|---|---|---|
|
|
Yeast extract | 2 | g | ||
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Peptone | 3 | g | ||
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Glucose | 5 | g | ||
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Acetic acid | 40 | ml | ||
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Ethanol | 30 | ml | ||
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Distilled water | 930 | ml | ||
| 1 The ethanol and acetic acid are sterilized separately by filtration (use filter resistant to these solvents), and added aseptically to the sterile medium. Acetobacter europaeus will not grow on plates containing 1-2% agar, and a double layer system has to be used. A layer of growth medium containing 0.5% agar (about 35 ml) is poured into a plate and allowed to solidify. A thin layer (5-10 ml) of the growth medium containing 1% agar is then poured on top. Carefully streak the plates, which are best incubated in closed containers (e.g. plastic boxes) at 30°C. | |||||