I am trying to regrow yeast that has been dried on filter paper. I have tried the following steps and have failed:
I have dabbed the filter paper on ypd agar plates and had growth but I failed to grow a colony from the plate in broth. I used ypd broth without supplements. should I incorporate amino acids such as leucine to get it to grow? And how much broth to use and what size flask? I put 50ml broth in 250ml flask at 250 rpm and 28-30 degrees.
Thank you in advance.
Undoubtedly YPD medium is enough. But I suppose you may try to grow yeasts from a plate in a stationary culture with 100-150ml of medium per 250ml flask (different aeration conditions). Maybe it will help...
I think you can try with Sabouraud-dextrose Agar and Sabouraud Broth. To make it safer, it is good to have chloraphenicol preadded to the medium. The medium volume you use seems perfectly right to me, just as well as the rotation speed. You can also try to cope with the temperature if you know the most optimal temperature for the yeasts you try to revive. It's not that simple to regrow yeasts from a waterless source.
Maybe start with a smaller overnight culture of 2 ml in a test tube and then sub inoculate that into a larger volume for proper growth. Best.
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