How do I use the 240 mL FastOrange broth bottles for sample enrichment?

The 240 mL FastOrange Broth bottles can be used for sample enrichment as long as quantities are scaled appropriately and solution is transferred using sterile technique to prevent contamination. It is recommended to aliquot the entire 240 mL broth bottle into individual enrichment containers at one time to minimize contamination. Unused enrichment containers can be stored at room temperature (max 25 °C) protected from light.

  1. Determine what enrichment containers to use. We recommend the following types of sterile enrichment containers for FastOrange enrichment:
  • T-75 cell culture flasks: Recommended for most enrichments as one-use containers. We suggest using these for standard 50 mL volume enrichments. 
  • Reusable glass bottles: Recommended as a cost-effective alternative if an autoclave is available
  • 50 mL Falcon/Eppendorf centrifuge tubes: Recommended for enriching bacteria growing in anaerobic environments such as Megasphaera, Pectinatus and L. acetotolerans* or  general purpose 25 mL volume enrichments. 
  • 15 mL tubes: Recommended for low-cost enrichments where the highest degree of sensitivity is not required. 

In general, use containers that will stand vertically to minimize air exposure when enriching using FastOrange B or FastOrange Brett Broth. When enriching using FastOrange Yeast or Wild Yeast Broth, find a container that can be laid flat on its side for maximum air exposure.

  1. Aliquot the FastOrange Broth into your enrichment containers. Carefully aliquot the desired quantity of solution as necessary until you have transferred your total required volume of broth (typically 40 – 50 mL) into each enrichment container based on graduations in the receiving container. Store the containers vertically at room temperature (max 25 °C), protected from light. 
  1. Add a volume of sample in a 1:1 ratio with the FastOrange Broth in one enrichment container using sterile technique. Transfer the total required volume of sample into the enrichment container (i.e., if you have a cell culture flask with 40 mL enrichment of FastOrange enrichment solution, pour in 40 mL of sample). Incubate according to the instructions of the specific enrichment broth. 

General Notes:

Store FastOrange Broth in the dark at room temperature (max 25 °C). If you’re using the broth to detect beer spoilers via color change only, protect from light during the sample enrichment. If you’re following up the enrichment with analysis via PCR, the enrichment container does not have to be protected from light during sample enrichment.

*For Megasphaera, Pectinatus and L. acetotolerans, please refer to the article on how to enrich for anerobic bacteria.