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OfflineSurfingmycelium
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Liquid culture issue
    #26957884 - 09/27/20 06:57 PM (3 years, 3 months ago)

Hey guys,

  I have this white stuff floating at the top of my liquid culture.  It just formed recently and it’s in both of my jars.  The jars were made weeks apart from each other.  One was injected with spores and the other had a piece of clean agar dropped in.  If I spin the magnetic bar around the stuff just stays at the top.  I’m guessing it’s contaminated?  It was doing so well for about two weeks.  Anyone ever see this?


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OfflineCocaineBuffet
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Re: Liquid culture issue [Re: Surfingmycelium]
    #26957889 - 09/27/20 07:00 PM (3 years, 3 months ago)

Since you have agar just test it and see. You would want to that anyways before adding it to grain anyways unless you wanted to only find out after all your grain turned to shit.


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InvisibleJosex
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Re: Liquid culture issue [Re: CocaineBuffet]
    #26957898 - 09/27/20 07:10 PM (3 years, 3 months ago)

It's totally normal for myc to start colonizing the surface once the colony has grown enough in the broth. We need pics to tell for sure.

Also, never inoculate an LC with spores, that's a sure fire way to fuck up. Spores are never clean.


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OfflineSurfingmycelium
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Re: Liquid culture issue [Re: Josex]
    #26958007 - 09/27/20 09:08 PM (3 years, 3 months ago)

Yeah I know.  I learned not to put spores to LC after the fact.  Still a newb.  This is agar that I put some of the spore liquid culture to about a week ago to see if it was contaminated.  This is before the floating white stuff showed up.



And here is the jar of the spore LC with the floating white stuff.



And here is the new jar I put a piece of clean agar too.  This is a completely different strain then the other and inoculated months apart from the other. 



When mixed it does not sink to the bottom.


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InvisibleJosex
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Re: Liquid culture issue [Re: Surfingmycelium]
    #26958318 - 09/28/20 05:10 AM (3 years, 3 months ago)

The pics are truly bad to tell for sure.


This can perfectly be cube myc. You'll know it's mold if it grows abnormally fast or changes color (sporulates). Sometimes, myc on the surface won't sink even if you swirl the jars, it's not usual but I've seen it happen.


If this is the test plate from the "spore to LC" jar, then I'd say it looks surprisingly clean. I don't see bacteria there, but it's way too desorganized and all over the place to tell for sure. Next time try squirting just a couple drops only.


As I said, the pic's really bad. At any rate, you can't tell if an LC is contaminated from a pic. Visual sings of contamination are things like liquid turning turbid and myc sinking to the bottom.

Sometimes a contaminated LC won't show visual signs of contamination.
The only way to know for sure is testing the LC on agar, and the agar looked ok to me, maybe try testing the LC on agar again but using just a drop or two.

I like my LC's to be very clear, so I make my broths at 0.2% strength using LME only. The more nutes you add the more difficult it gets to detect contamination.

Look up the first link in my sig, that's how I make LC's. You're going to find lots of interesting info there.


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OfflineSurfingmycelium
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Re: Liquid culture issue [Re: Josex]
    #26958566 - 09/28/20 10:05 AM (3 years, 3 months ago)

Here is a better pic.  I was able to get a better shot with the sunlight coming in this morning



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InvisibleJosex
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Re: Liquid culture issue [Re: Surfingmycelium]
    #26958572 - 09/28/20 10:09 AM (3 years, 3 months ago)

That's cube myc man. :thumbup:


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OfflineSurfingmycelium
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Re: Liquid culture issue [Re: Josex]
    #26958631 - 09/28/20 11:02 AM (3 years, 3 months ago)

Sweet, thanks!  Now I just have to figure out why it isn’t colonizing my grains.  Lol.  Been two weeks and nothing.


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InvisibleJosex
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Re: Liquid culture issue [Re: Surfingmycelium]
    #26958768 - 09/28/20 12:45 PM (3 years, 3 months ago)

Well there's what appears to be healthy myc on the test plate. You need a really bacterial LC for it to not leap off on the grain, so bacterial it would have been pretty obvious on the test plate.
So I would not discard the possibility of bad grain. I had something similar happen once with oats, the LC wouldn't leap off. What grains are you using? How are you prepping them?


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OfflineSurfingmycelium
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Re: Liquid culture issue [Re: Josex]
    #26959560 - 09/28/20 11:18 PM (3 years, 3 months ago)

I got them off a reputable site in the “Midwest.”  It’s rye grain.


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