The enrichment process for acetic acid bacteria is similar to that of lactic acid bacteria, but optimal growth conditions vary. Unlike lactic acid bacteria, acetic acid bacteria grow best in aerobic (oxygen present) environments, so it is important to maximize surface area as much as possible during the enrichment process. FastOrange B solution provides a limit of detection of 1 cell/sample in 48 hours when followed by qPCR. A volume of 50 mL is recommended for sensitive detection. Less volume of FastOrange B solution may be used, but this will result in decreased sensitivity.
Using the 50 mL FastOrange B Enrichment Bottles:
- Add 50 mL of your sample to a 50 mL FastOrange B Enrichment Bottle using sterile technique and incubate it for 48 hours at room temperature (25 °C +/- 2 °C). Leave the bottle flat on its side to maximize oxygen exposure and vent the sample to avoid pressure buildup.
Using the 240 mL FastOrange B Broth:
- Aliquot your entire 240 mL FastOrange B Broth from the glass bottle into individual enrichment containers with your chosen volume. We recommend 50 mL for sensitive detection. Use sterile technique to transfer the solution.
- Add your sample in a 1:1 ratio to one enrichment container, again using sterile technique. Incubate the enrichment container for 48 hours at room temperature (25 °C +/- 2 °C). Leave the bottle flat on its side and vent the sample to avoid pressure buildup.
After your sample has been enriched for 48 hours, proceed with analysis by qPCR.