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OnlineBakedbeings
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Re: Questions for first attempt [Re: msj]
    #27342701 - 06/10/21 10:46 AM (8 days, 3 hours ago)

do those condiment cups arrive sterile? i guess at this point we hope so


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Re: Questions for first attempt [Re: Bakedbeings]
    #27346252 - 06/13/21 02:21 AM (5 days, 11 hours ago)

Quote:

Bakedbeings said:
do those condiment cups arrive sterile? i guess at this point we hope so




I've seen someone use them before who seems to have good results, and so far so good I'm hoping.  I'm assuming at the three day mark I should see nearly nothing, which is what I have. 

I've never grown mycelium on agar before, though, so to be fair I have no idea what it looks like.  From what I can tell, there are some teensy tiny itty bitty dark purple dots in some of the cultures, which look to me to be smaller clumps of the spores because they're the same color as the clumps in the syringe were. 

Also, near some of these spots (and also independent of any visible spots in some dishes), there is the faintest suggestion of what almost looks like a snowflake pattern.  This is only in a few of the dishes so far.  It's barely visible to the naked eye right now, and it looks like filaments reaching outward from this one center point, and the filaments are pretty strikingly white in contrast to the color of the agar.  The agar I used is a light to medium golden amber color, which I made with 10g agar-agar and 10g LME to just shy of 500ml distilled water.  The dishes have been held at a steady 80-82 degrees for the past three days since inoculation.

Is this a good sign?  What do you specifically notice first that lets you know you've got mycelium and not something else growing?  Or is it more a case of "well it could be anything" until it's bigger?  I've googled but haven't found many good pictures of cube mycelium at 3 days old.

My sterile petri dishes finally got here so I made another batch tonight, but does it sound like the first ones are on track?  Trying hard to hedge my bets so this endeavor is successful.


Edited by msj (06/15/21 12:14 AM)


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Re: Questions for first attempt [Re: msj]
    #27346786 - 06/13/21 02:49 PM (4 days, 23 hours ago)

sounds like you can afford to be optimistic but at this point only time will tell


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Re: Questions for first attempt [Re: Bakedbeings]
    #27348079 - 06/14/21 05:42 PM (3 days, 20 hours ago)

I'm gonna take pictures when it's a little easier to see because I really should have colored my agar plates (I did not realize how much I would want the contrast to see better).  However, after taking a quick look today...

I HAVE MYCELIUM BABIES!  They're so cute!  The condiment cups seem to have worked fairly well. 

I'm considering sacrificing one of the 25 I have by opening it to the open air and seeing how quickly it becomes contaminated just so I have some basis for comparison about growth rates and can actually see what pops up from it being exposed to the open air in my house.  I figure if I grow the contaminants on purpose, I would know what they'll typically look like for my environment.

I've heard through my research that agar made with 7.5 grams LME, 10g agar, 500ml water, and .25g powdered nutritional yeast is good for encouraging rhizomorphic growth when you transfer to another agar plate.  The reasoning behind it is that the agar contains less LME so the mycelium has to "reach" for nutrients (and something about the nutritional yeast but tbh that part wasn't well explained).  Going to experiment with my agar formulations a bit, so any advice there is welcome.

Editing to add to this because I looked and one (but thankfully only one) particular cup that I had issues with while pouring it appears very different from the others.  I'm pretty sure I need to toss this one.  Whatever this is, it seems to grow a lot faster, it grows up above the agar surface, it doesn't appear to grow from a central point like the other cups are exhibiting, and it's much, much less dense than I would expect mycelium to be.





Edited by msj (06/15/21 12:08 AM)


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Re: Questions for first attempt [Re: msj]
    #27348907 - 06/15/21 10:03 AM (3 days, 4 hours ago)

yea i was gonna say dont worry about contamming your shit on purpose, youll get plenty of experience without trying :smile:

looks to me like cobweb mold


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Re: Questions for first attempt [Re: Bakedbeings]
    #27350119 - 06/16/21 03:31 AM (2 days, 10 hours ago)

Hahahaha you are so right, turns out.  I think if you don't sterilize these somehow it can go any kinda way. 

Out of the 25 I did, roughly one third are looking like healthy mycelium, roughly one third are contaminated with something or other, and another third have nothing at all growing.  The biggest issue I have so far is telling what's what when it's so small.  The cobweb sort of fools you until you start to see the little gray specks but by the time you see gray specks in a condiment cup the whole thing is practically overtaken.

I know I need to keep cleaning up the mycelium by doing multiple transfers until I'm certain it's totally clean and pretty.  But once I get to that point, from my understanding, I have some options about how I want to proceed with inoculating my grain jars.  So here's my question.

If you were me doing this all for the first time, would you attempt to make a liquid culture jar from your clean mycelium once you have it in order to inoculate your grain jars?  Or would you inoculate your jars with mycelium on agar directly?  Is there really much of a difference since they're both live mycelium?  The more I read about liquid culture the more I want to make a gigantic jar full of it.


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Re: Questions for first attempt [Re: msj]
    #27350122 - 06/16/21 03:36 AM (2 days, 10 hours ago)

Quote:

msj said:
If you were me doing this all for the first time, would you attempt to make a liquid culture jar from your clean mycelium once you have it in order to inoculate your grain jars?  Or would you inoculate your jars with mycelium on agar directly?  Is there really much of a difference since they're both live mycelium?




Mega difference :wink:
LC cuts down colonization times in half.
If you pour some LC to grain jars and distribute it carefully,
imagine how many more inoculation points you have instead of an agar piece.

              :cookiemonster:


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Re: Questions for first attempt [Re: Goatrider]
    #27350862 - 06/16/21 04:40 PM (1 day, 21 hours ago)

Thank you for the clarification.  I'm gonna attempt the LC at some point soon, I think it's a great way for me to go.  In the meantime, found this on one of my other petri dishes:



After googling I'm pretty sure that's trich if I'm not mistaken.  Would you say to just go ahead and toss the whole plate or is it worth opening if there's mycelium growing on the other side?  Kind of hesitant to even pop the lid, thinking of throwing it straight into the outdoor trash can.


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Re: Questions for first attempt [Re: msj]
    #27350889 - 06/16/21 04:57 PM (1 day, 21 hours ago)

That's some aspergillus or penicillium.
You can transfer, no problem.
Just open the lid as much as needed to reach the transfer point.
If it's worth, do it.

              :cookiemonster:


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