Enriching samples for Diastaticus requires a 48-hour enrichment using the FastOrange Wild Yeast solution, which provides a limit of detection of 1 cell/sample in 48 hours when followed by qPCR. A volume of 40 mL is recommended for sensitive detection. Less volume of FastOrange Wild Yeast solution may be used, but this will result in decreased sensitivity. Diastaticus grows in aerobic environments, so lay your enrichment containers on the side to maximize air exposure when possible. All enrichment solution and sample volume should be transferred utilizing sterile technique.
Using the 40 mL FastOrange Wild Yeast Enrichment Bottles:
- Add 40 mL of your sample to a 40 mL FastOrange Wild Yeast Enrichment Bottle and incubate for 48 hours at room temperature (25 °C +/- 2 °C). Lay the bottle flat on its side and vent the sample to avoid pressure buildup.
Using the 240 mL FastOrange Wild Yeast Broth:
- Aliquot your entire 240 mL FastOrange Wild Yeast 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. Aliquoted enrichment containers can be stored at room temperature (max 25 °C), protected from light.
- 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 container 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.