Inoculating and Culturing the Dregs of Avery 15

Culturing Yeast From Avery’s 15th Anniversary Beer

A few days prior to inoculating plates, I removed the special foil (and glue used to hold the foil) from the top and underside of an Avery 15 bottle cap by thoroughly cleaning and scrubbing with warm soapy water. After rinsing and drying the bottle, the neck of the bottle including the cap was soaked in sanitizing solution and the bottle was sat up right to let the yeast cells re-collect in the bottom. Avery’s 15th Anniversary beer is one of the first 100% Brettanomyces fermented beers and was produced with a unique strain not available to homebrewers. The strain they used is refereed to as “Drie“, as it was cultured from a Drie Fonteinen gueuze bottle by the Brewing-Science Institute.

Using the plates poured a day earlier, I pulled out 6 MYPG w/CaCO3 and 6 standard MYPG plates. Using sterile lab techniques throughout the entire process, I opened the bottle, flamed the lip with a lit Bunsen burner and poured the beer into a glass. I left approximately 90 ml of dregs in the bottle (too much only needed about 20 ml) and thoroughly roused the dregs giving a good mixing. The dregs were then aseptically poured into five, 20 ml pre-sterilized (autoclaved) vials with screw tops.

  • 3 of each agar plate (6 total) were inoculated with .1 ml of dregs pipetted onto the plate then dispersed via glass beads
  • 2 of each agar plate (4 total) were inoculated by looping cell and streaking for single colonies
  • 1 of each agar plate (2 total) had 1 ml pipetted onto the plate then dispersed via glass beads (This is too much inoculate, .1 ml is more then enough!)

After the inoculate absorbed into the plates the plates were then placed upside down, labeled accordingly, and placed in a culturing chamber set at 28° Celsius.

I will leave them for an initial 48 hours and keep observing till cell growth has occurred and I’m able to collect single colonies for species identification and propagation. I will post photos of the plates to show the growth and any possible zones of clearing on the MYPG w/CaCO3 inoculated plates if acids are produced that quickly.

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