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Offlinefinalreview
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Registered: 12/01/13
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Last seen: 10 years, 2 months
Agar scenario
    #19243915 - 12/08/13 08:11 AM (10 years, 2 months ago)

Hypothetically what would happen if an exited young mycologist were to innouclate a stack of agar plates while they were still cooling? Let's say he took the bottle of agar out of the PC, placed it into his prepped glovebox, then poured 15 plates. Then after filling the last plate he started syringe inoculating the plates starting with the first poured. Would this procedure fail in creating myc? Also what if some of the black spores in the syringe were released with the liquid onto a plate?


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OfflineCliftonGK1
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Re: Agar scenario [Re: finalreview]
    #19243972 - 12/08/13 08:34 AM (10 years, 2 months ago)

The spores are what will grow into mycelium, so make sure to agitate the syringe before inoculation to assure that they are evenly dispersed.

As for the heat, how hot are we talking here?  The plates had already set, right?  Agar plates don't need to be poured very thick and they tend to cool quickly, so it's not likely that you inactivated the spores by inoculating onto warm media.


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Invisiblemycomattie
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Re: Agar scenario [Re: CliftonGK1]
    #19244279 - 12/08/13 10:12 AM (10 years, 2 months ago)

If the agar temp is > 106 F the spores would simply die - that's the thermal death temp for cubensis spores.


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InvisibleSpitballJedi
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Re: Agar scenario [Re: finalreview]
    #19244354 - 12/08/13 10:35 AM (10 years, 2 months ago)

The purpose of agar plates is to create a flat, single plane for your culture to grow out on. The flat surface makes it easier to spot contams and sectors because they are growing, practically, in 2 dimensions.

If your agar is too soft, or still liquid, then the purpose is defeated. The culture would have the opportunity to grow in the vertical dimension and contams could more easily be hidden underneath.


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InvisiblePastywhyteMDiscord
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Re: Agar scenario [Re: SpitballJedi]
    #19244484 - 12/08/13 11:08 AM (10 years, 2 months ago)

Quote:

SpitballJedi said:
If your agar is too soft, or still liquid, then the purpose is defeated. The culture would have the opportunity to grow in the vertical dimension and contams could more easily be hidden underneath.



Actually this is a technique that is good for cleaning up cultures, I do it all the time. The mushroom mycelium can push though the agar media and will always try to grow to the surface, while most molds and bacteria cannot. The result is that the act of growing through the mycelium strips out the contams leaving clean mycelium on the surface.

An excellent tek here on the boards utilizes this principle, has helped me clean up many a contamed culture.

http://www.shroomery.org/forums/showflat.php/Number/2823255#2823255

I just skip the peroxide. It works very well though. Usually I don't wait for it to grow off the sample as sometimes bacteria can worm its way through the sandwich. I usually just tease of some of the clean mycelium with loop as it comes through the top.


Edited by Pastywhyte (12/08/13 11:17 AM)


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OfflineCliftonGK1
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Re: Agar scenario [Re: Pastywhyte]
    #19244810 - 12/08/13 12:42 PM (10 years, 2 months ago)

Quote:

Pastywhyte said:
Actually this is a technique that is good for cleaning up cultures, I do it all the time. The mushroom mycelium can push though the agar media and will always try to grow to the surface, while most molds and bacteria cannot. The result is that the act of growing through the mycelium strips out the contams leaving clean mycelium on the surface.





Sticking this one in the memory vault.  Thank you!


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Offlinefinalreview
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Re: Agar scenario [Re: CliftonGK1]
    #19245620 - 12/08/13 03:38 PM (10 years, 2 months ago)

Thanks for answering!


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