| | Glucose | 40 | g | | | Peptone | 10 | g | | | Yeast extract | 10 | g | | | Citric acid·H2O | 1.5 | g | | | Na2HPO4·2H2O | 3.38 | g | | | Glacial acetic acid | 10 | ml | | | Absolute ethanol | 10 | ml | | | Distilled water | 980 | ml | | | pH 3.8 (pH is not adjusted) | | | | | 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 sterilized either by filtration (use PTFE membrane filters) or they may be autoclaved in completely closed screw cap glass bottles sealed with a PTFE coated septum. The preparation of solid media employs a double layer system. A layer of growth medium containing 0.5% agar (about 20 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 and incubate in closed containers to keep the humidity high (>90%). |
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