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Offlineadrenalytic
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First Time Agar. What's going on here?
    #25837265 - 02/25/19 07:11 PM (2 years, 7 days ago)

Hello,

I'm still very new to the mushroom cultivation gig and I've recently begun dabbling in the agar business.  I took some tissue samples of a mushroom that I liked and am attempting to clone it in agar.  The problem is, I can't tell if some of the stuff is mycelium or not.  I know mycelium is fuzzy and ropey looking on the surface but what does it look like under agar? 

Here are some pics.

This is the top view:



And from the bottom:



So I'm betting the fuzzy white one in the top left corner is mycelium.  Is the slimy glistening looking one just underneath the agar or is that a contam?

If it's contam, what is the best removal method so as not to disturb the healthy myc?  Again, I am just starting to learn agar so I've yet to read everything about it but I have not seen a decent tek outlining how to remove contams yet.

Thanks.


Edited by adrenalytic (02/25/19 07:17 PM)


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OfflinePsilocyBen17
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Re: First Time Agar. What's going on here? [Re: adrenalytic]
    #25837292 - 02/25/19 07:25 PM (2 years, 7 days ago)

looks like a lot of bacteria in there...what are you doing to keep things sterile? ive never seen a dish wrapped with micropore tape


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Offlineadrenalytic
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Re: First Time Agar. What's going on here? [Re: PsilocyBen17]
    #25837303 - 02/25/19 07:31 PM (2 years, 7 days ago)

I use a SAB for transfer.  I read in a tek to use the tape because it protects from contam and allows for FAE.

Edit: and I used 70% iso to clean the fruit before I cut it open to collect tissue.

Edit 2: I have 3 other dishes.  One looks like it's all slimy shit on the three samples I dropped in.  The other two look like nice healthy myc.


Edited by adrenalytic (02/25/19 07:37 PM)


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OfflineNo-shiitakeman
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Re: First Time Agar. What's going on here? [Re: adrenalytic]
    #25837427 - 02/25/19 08:27 PM (2 years, 7 days ago)

It's near impossible to clean up an agar dish.

It's standard practice  to remove a small piece of the healthy mycelium, to a clean agar dish using a flame sterilized scalpel.

Also when I take a take a tissue culture to clone I wipe the outside of the shroom with an alcohol wipe, let it dry then tear it in half from the base of the stem to the cap. Then break the cap in half. Tearing  instead of cutting keeps from forcing contaminants that are on the surface of the mushroom into clean tissue.



The top left one is the only one that doesn't look horribly contaminated. Hopefully in a few days the mycelium in that will grow fast enough for you to move a small sample to a clean dish. It's still contaminated so Youll have to be very careful when doing it. It will be a project.


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OfflineBeefSupremeJr
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Re: First Time Agar. What's going on here? [Re: No-shiitakeman]
    #25837548 - 02/25/19 09:11 PM (2 years, 7 days ago)

Looks like you got some growth on the top but and maybe even the bottom black bit.  I’d say go ahead and let it grow a while and take some transfers from there carefully. 

That yellow spot I am told is  typically staph.


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Offlineadrenalytic
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Re: First Time Agar. What's going on here? [Re: adrenalytic]
    #25837568 - 02/25/19 09:20 PM (2 years, 7 days ago)

Thanks for the replies so far.  Just so there's no confusion, the black you see is actually just the bruised tissue on the shroom from collection.  The sample I collected it from was sitting in the fridge for 5 days before I collected it so it bruised very easily.


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OfflineBeefSupremeJr
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Re: First Time Agar. What's going on here? [Re: adrenalytic]
    #25837575 - 02/25/19 09:23 PM (2 years, 7 days ago)

Quote:

adrenalytic said:
Thanks for the replies so far.  Just so there's no confusion, the black you see is actually just the bruised tissue on the shroom from collection.  The sample I collected it from was sitting in the fridge for 5 days before I collected it so it bruised very easily.





I know :randysavage:

Next time cut your samples as your box is already setup to work.  Fridge’s are gross.


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Offlineadrenalytic
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Re: First Time Agar. What's going on here? [Re: BeefSupremeJr]
    #25837714 - 02/25/19 10:10 PM (2 years, 7 days ago)

Quote:

BeefSupremeJr said:
Quote:

adrenalytic said:
Thanks for the replies so far.  Just so there's no confusion, the black you see is actually just the bruised tissue on the shroom from collection.  The sample I collected it from was sitting in the fridge for 5 days before I collected it so it bruised very easily.





I know :randysavage:

Next time cut your samples as your box is already setup to work.  Fridge’s are gross.




I didn't have a choice.  I didn't have agar plates for the mushie I wanted to clone. Had to wait for delivery.


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OfflineBeefSupremeJr
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Re: First Time Agar. What's going on here? [Re: adrenalytic]
    #25837716 - 02/25/19 10:11 PM (2 years, 7 days ago)

Next time :randysavage:


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OfflineMr. Funguy
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Re: First Time Agar. What's going on here? [Re: BeefSupremeJr]
    #25837915 - 02/25/19 11:37 PM (2 years, 7 days ago)

Looks like yellow slime mold, give it a week see if it strands out. Lol


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OfflinePsilocyBen17
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Re: First Time Agar. What's going on here? [Re: Mr. Funguy]
    #25839499 - 02/26/19 06:42 PM (2 years, 6 days ago)

i wouldnt let your samples sit in the fridge. i would harvest them and clone asap. think of all the rot and nastiness they encounter in a fridge. i would bet thst is the source of your problems


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