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OfflineHikeadellic
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Registered: 08/31/20
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Spores not germinating on agar?
    #26974599 - 10/07/20 10:06 PM (3 years, 3 months ago)

Hey all, spores were placed on agar plates over 2 weeks ago. The plates are very small, about a little bigger then a quarter coin. No mycelium growth has been observed. They were transferred to the plates by scraping them from the tin foil print to the plate with a small, alcohol and flame sterilized metal scrappy thing in a SAB. Is something wrong? Is more time needed? Do the spores need to be totally submerged in agar? Are more practical methods of spore germination available?


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InvisibleRhizomorph
Psychedelic Researcher
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Registered: 04/24/20
Posts: 785
Re: Spores not germinating on agar? [Re: Hikeadellic]
    #26974659 - 10/07/20 11:08 PM (3 years, 3 months ago)

Did you cool off the flame sterilized tool before scraping the spores with it?

A single speck of spores has thousands of individual spores in it. Just 2 spores on a nutrient-dense medium like agar would be all they need to germinate. If you made your agar properly and you didn't somehow kill the spores/render them nonviable they should have germinated.

Do the plates have gas exchange? Pictures may help.


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Offlineverum subsequentis
seeker of truth
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Registered: 03/22/16
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Re: Spores not germinating on agar? [Re: Rhizomorph]
    #26974725 - 10/08/20 12:05 AM (3 years, 3 months ago)

Spores don't need gas exchange to germ because they aren't breathing.

Some times spores take their sweet time. Some other times spores never germ because they are old and dead. Some folks prefer to make soft agar as to allow the spores to gather moisture and believe it helps. I don't bother but i would if i ran into some spores that i really wanted to germ and couldn't via the usual methods.

Some folks believe in hydrating spores in sterile water to help with germ. I haven't needed to do that so i can't speak to it.


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OfflineMrTheseThings
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Registered: 06/21/20
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Re: Spores not germinating on agar? [Re: verum subsequentis]
    #26974734 - 10/08/20 12:14 AM (3 years, 3 months ago)

Sometimes it takes me a few tries . Imo it's worth another shot. I have had this happen more than once.


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OfflineHikeadellic
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Registered: 08/31/20
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Re: Spores not germinating on agar? [Re: MrTheseThings]
    #26975143 - 10/08/20 09:11 AM (3 years, 3 months ago)

To my knowledge, the print was about a month or so old. I did not bring the scrape tool to red hot temps. I gave it a minute to air cool prior to use.  I just wiped it with 70% iso and made a few passes with a lighter. It was a BRF-Agar recipe that has worked in the past. I've never seen plates this tiny so I thought maybe after wrapping them that gas exchange may be an issue. Never came across any teks mentioning gas exchange on plates either so who knows. I guess Ill just wait it out and see what happens.


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InvisibleRhizomorph
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Registered: 04/24/20
Posts: 785
Re: Spores not germinating on agar? [Re: verum subsequentis]
    #26975217 - 10/08/20 10:16 AM (3 years, 3 months ago)

Thanks verum! I didn't know that. Now I do :grin:


--------------------

:cookiemonster: Major Issues in the Psychedelic Movement: Why 'Psychedelic People' and the Psychedelic Movement Sucks:elmo:

:awesomenod: Easiest No-Pour Agar Method: Alien's Holy Grail No pour Agar unmodified containers:awesomenod:


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