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I made about a dozen plates over the weekend using a tryptone/yeast-extract medium. All plates and medium were autoclaved @ 250 F for 30 min. With a sterile innoculation loop, I swiped directly from the spore print to the agar. Innoculated plates are being stored in darkness around 80 F. I checked them this AM and am not seeing any indication of growth. I know that I am probably being prematurely worried, but I have a couple of questions after the fact. Should I have hydrated the spores prior to the agar transfer? How long should I wait before my attempt is ruled a failure? I poured my plates a little thick (~ 3 mm), will the mycellium penetrate the agar, or just grow across the surface.
depending on how old the spres are it could take 5-7 days. they will rehydrate on the agar. and it grows across the agar. 250 for 30 is a little long, 15 minutes should be it.
I would wait at least ten days before you give up on the plates. but its not a bad thing to pour your plates a little think (my cat's plates are usually around 4 mm) just because the mycelium will have a ready source of water when you drop your agar into your grain. If your agar is too thin it will dry out before the mycelium really gets a good grip on the grain and can sustain itself.
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