Acetobacter Xylinum Culture
General guidelines on how to grow up a culture of Acetobacter xylinum, ATCC strain 53582 .
Preparation of Acetobacter Media
To prepare ∼500 ml of liquid Acetobacter media, add the following:
- Glucose - 10 g
- Peptone - 2.5 g
- Yeast extract - 2.5 g
- Na2HPO4 - 1.35 g
- Citric acid - 0.75 g
- Distilled water - 500 ml
- If you are making plates, use the same protocol but add 7.5 g of agar.
- Prepare media as outlined
- Autoclave to sterilize media.
- Streak/inoculate Acetobacter onto plates or in media.
- Incubate cells at 26°C for 2-3 days.
- If using a freeze dried source of Acetobacter (ex. ATCC shipment), growth may take up to 4 days.
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- The growth of Acetobacter does not give a cloudy appearance in the media, the media will remain transparent to slightly translucent in appearance.
- The growth of Acetobacter is accompanied by the formation of a thick cellulose matrix within the media that must be removed before cells can be pelleted for a miniprep procedure. Simply vortex briefly to break up the cellulose into chunks and remove the cellulose chunks from the media with a pipette while carefully avoiding the removal of cells.
- Acetobacter will grow well at room temperature in aerobic conditions.
- For information on the growth conditions of other Acetobacter strains, please visit 
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