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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Agar culture question
    #25804543 -

http://

So I have this culture here of panaeolus cyan Hawaii inoculated 2/4 on MEA agar. This is my first time doing agar and my contam rate was about half so far. I have some viable cultures growing though. This one the spore just germinated a couple days ago but there is some contam in the dish. Should I transfer and rescue this mycelium now before the contam gets to it? Or is it too early.

Also should note that when I used the syringe it blasted so much solution onto my dish maybe a cc each (wasn’t on purpose, just gotta practice syringe technique lol) so there is standing water that has a weird color to it. You can see it in the picture if you look close enough.


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25804576 -

Not an expert, but a cc each is going to contain all the contams.  I’m not sure you could transfer away through a layer of unclean water.

All hail the myco god who can.  Teach me, o master.

Edit: A better idea is to sterilize a shot glass in your PC, and inject some into it.  Then use a loop to inoculate your dish.


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Edited by Ep1429 (02/10/19 11:23 PM)

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Re: Agar culture question [Re: Ep1429]
    #25804606 -

Damn the shot glass would’ve been much better. I stabbed my finger by accident too lol but at least it was sterile :shrug: 

The contam seems to be contained to the edges/that one yellow spot on there. But the white looks good, I might cut that out and transfer, I don’t have too many plates to work with, and three species to work on. The agar is more practice for cloning anyway. As long as I can nocc some grain :mushroom2:


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25804635 -

Funnguiy said:
Damn the shot glass would’ve been much better. I stabbed my finger by accident too lol but at least it was sterile :shrug: 



OH damn, you got it now. Spores in the blood is no joke. they are now making their way to your brain and spinal cord. You'll never be the same again. It's ok though, many of us are infected with the magic seeds and still function pretty well.

I just flame the needle and then squeeze a tiny bit out (to cool the needle) and the carefully push until a drip is dangling of the end of the needle. If some shoots out onto the table it's no big deal but i try not to waste any. i then grab the dangling drip with my (flamed) loop and streak the plate.

There is no such thing as "too small to transfer" unless you are waiting for the culture to out run something. If you have standing water covering the surface of the agar you'll almost definitely not get a clean culture on your first transfer but you very well may on the second or third. If their is no contam touching the culture or touching any water that is touching the culture than you might be just fine.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Agar culture question [Re: verum subsequentis]
    #25804649 -

I always go to grain from syringe and then section out good growth to plates and then cleanbfrom there but IMA strange bastard


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CA weed @ all times unless specified
currently this high
pebbles suck
its turtles all the way down

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Re: Agar culture question [Re: psilly the kid]
    #25804658 -

are you saying colonized grain to plates or cloning tissue to plates.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Agar culture question [Re: verum subsequentis]
    #25804660 -

Both......


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CA weed @ all times unless specified
currently this high
pebbles suck
its turtles all the way down

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Re: Agar culture question [Re: psilly the kid]
    #25804689 -

Lolll no it wasn’t on the syringe yet I was getting the needle guard off a little too enthusiastically and it jabbed me rat bastard.

Yeah, I have another panaeolus plate but it’s the only one that hasn’t germinated yet however is clean. I’m doing a test, I have three species, and plan on six rye jars. 2 jars for each species, one nocced hopefully with agar and the other I think syringe. I think I’ll cut the culture out tomorrow and roll with that.


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25804693 -

Why risk your jar do all agar


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CA weed @ all times unless specified
currently this high
pebbles suck
its turtles all the way down

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Re: Agar culture question [Re: psilly the kid]
    #25804764 -

We’ll see how these agar cultures turn out, but I might just do all agar idk.


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25806113 -

Also, I have plates un inoculated with little spots of contam. Can’t I just cut out the isolated contamination and theoretically be able to use it?


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25806158 -

theoretically sure. Chances of success. .00000000000000006%


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Agar culture question [Re: verum subsequentis]
    #25806180 -

You could let it get just a little bigger if you keep a *real* close eye on it just to make sure you are looking at myc germination.

But look up at 11oclock - see that yellow blob of bacteria? There is also a thin layer of bacteria slime following the edge of the plate from like 9oclock to 2 o'clock on the plate. You don't want it to get too close to that.

I know you said you used a lot of spore solution, see how the bacteria formed along the edge where the liquid gathered?
Next time you inoculate with spore solution, use only a single drop in the center, and then use a flame sterilized inoculation loop to swipe the spores in a zig zag pattern across the plate. This way you'll be able to follow where germination should be occurring.


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Amateur Mycologists United
AMU Q&A

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Re: Agar culture question [Re: psilly the kid]
    #25806182 -

psilly the kid said:
I always go to grain from syringe and then section out good growth to plates and then cleanbfrom there but IMA strange bastard



I would really go the other way around man. Once bacteria/mold have intertwined with the mycelium you aren't likely to get them apart on agar IME


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Amateur Mycologists United
AMU Q&A

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Re: Agar culture question [Re: mushpunx]
    #25806880 -

Yeah, I don’t have much of a choice unless I try to nocc another plate from scratch, which I’d rather not. I’m 99.9% sure that white spot is mycelium. It has doubled since I took that picture and it rhizomorphic.
I’m letting it go tonight and tomorrow, and I’ll re evaluate tomorrow. Will probably transfer tho before the contam gets too prominent in the plate.


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25807860 -

i'd do it as soon as possible


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Agar culture question [Re: verum subsequentis]
    #25808329 -

Yea get that ASAP the bacteria slime is right above it!


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Amateur Mycologists United
AMU Q&A

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Re: Agar culture question [Re: mushpunx]
    #25808386 -

Just finished. I transferred two pieces from that white spot to two separate other plates, to hopefully increase the odds of a viable culture. I’m much more satisfied with my sterile technique this time around though.

I had about 6 plates un inoculated but each had a small either yellow or orangish dot, so I cut them all out and put in a cup with iso after all mushroom business was done. Idk if it will work, but I would’ve just tossed them anyways so I’ll see what happens. This is the learning part haha


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“Time flies. It’s up to you to be the navigator.”

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Re: Agar culture question [Re: Funnguiy]
    #25808659 -

Did you pour the plates or order pre poured? somebody messed up.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: Agar culture question [Re: verum subsequentis]
    #25808783 -

:whathesaid:


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Amateur Mycologists United
AMU Q&A

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Shop: Original Sensible Seeds Autoflowering Cannabis Seeds   Mushroom-Hut Liquid Cultures   Myyco.com Isolated Cubensis Liquid Culture For Sale   MagicBag.co All-In-One Bags That Don't Suck   Sporeworks.EU Spores for European Microscopy   Unfolding Nature Unfolding Nature: Being in the Implicate Order   North Spore Injection Grain Bag

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