How do I enrich for Brettanomyces?

FastOrange Brett solution provides a limit of detection of 1 cell/sample in 72 hours when followed by qPCR. A volume of 40 mL is recommended for sensitive detection. Less volume of FastOrange Brett solution may be used, but this will result in decreased sensitivity. Brettanomyces grows best in anaerobic (oxygen devoid) environments, so it is important to minimize the surface area as much as possible during the enrichment process.

Using the 40 mL FastOrange Brett Enrichment Bottles:

  1. Add 40 mL of your sample to the 40 mL FastOrange Brett Enrichment Bottle using sterile technique and incubate it for 72 hours at room temperature (25 °C +/- 2 °C). Leave the bottle upright to minimize oxygen exposure and vent the sample to avoid pressure buildup.

Using the 240 mL FastOrange Brett Broth:

  1. Aliquot your entire 240 mL FastOrange B Broth from the glass bottle into individual enrichment containers with your chosen volume. We recommend 40 mL for sensitive detection. Use sterile technique to transfer the solution. 
  2. 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 72 hours, proceed with analysis by qPCR.