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OfflineMyco816
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Is this LC ready for grain?
    #28772891 - 05/13/24 11:15 AM (1 month, 13 days ago)

I created these two jars on 4/29 from JMF agar that had been transferred twice. The jar on the left, I used the poke technique. The jar on the right, I dropped in a small wedge of agar.

The jar on the left has always been kind of cloudy - I wasn't sure if that's indicative of a problem, or if that's just the way the mycelium develops with the poke technique? The little clumps in the left jar formed in just the last couple days.

Are either/both these jars good to inoculate grain? Or is there an advantage to letting the myc colonize in the jars longer?

(I know you're generally supposed to test on agar first, although these came from clean agar plates so not sure if that's necessary?)


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Re: Is this LC ready for grain? [Re: Myco816] * 1
    #28772894 - 05/13/24 11:22 AM (1 month, 13 days ago)

:shrug: did u test it


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OfflineMyco816
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Re: Is this LC ready for grain? [Re: tripdawg420]
    #28772899 - 05/13/24 11:25 AM (1 month, 13 days ago)

No, I haven’t tested it yet.
Quote:

tripdawg420 said:
:shrug: did u test it




No, I haven’t tested it. I was planning not to open the lid to test until it was “done” brewing. (I don’t have a syringe port on the lid, only an SFD.)

Just not sure if these are done, or if I should wait until there is more myc in the jar before testing/inoculating?

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OfflineGorguss
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Re: Is this LC ready for grain? [Re: Myco816] * 1
    #28772903 - 05/13/24 11:29 AM (1 month, 13 days ago)

Dropping it on some agar is the way to determine if its ready or not. I myself always skip it and I have imo, higher contam rates than normal.

Test on agar


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Onlinehamloaf
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Re: Is this LC ready for grain? [Re: Myco816] * 3
    #28772916 - 05/13/24 11:38 AM (1 month, 13 days ago)

Quote:

Myco816 said:
I created these two jars on 4/29 from JMF agar that had been transferred twice. The jar on the left, I used the poke technique. The jar on the right, I dropped in a small wedge of agar.

The jar on the left has always been kind of cloudy - I wasn't sure if that's indicative of a problem, or if that's just the way the mycelium develops with the poke technique? The little clumps in the left jar formed in just the last couple days.

Are either/both these jars good to inoculate grain? Or is there an advantage to letting the myc colonize in the jars longer?

(I know you're generally supposed to test on agar first, although these came from clean agar plates so not sure if that's necessary?)





Yes, Cloudy-ness of LC is indicative of a contaminated LC, that is correct.  Do not expand any further with the jar on the left.  Also, inoculating LC with agar is the correct operating procedure. 

LC is an all or nothing gamble.  With that being said, until you are confident in your liquid culturing techniques, testing your LC on agar would be your safest bet before sending it to sterilized grains.


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InvisibleYuggoth
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Re: Is this LC ready for grain? [Re: hamloaf]
    #28773253 - 05/13/24 04:56 PM (1 month, 13 days ago)

> The jar on the left has always been kind of cloudy...

ALWAYS cloudy, like, from the time it came out of the PC?

I have made LC that was a little cloudy when freshly sterilized, due to ingredients. But if it was clear and it turned cloudy after inoculation, that is bad news.


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OfflineMyco816
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Re: Is this LC ready for grain? [Re: Yuggoth]
    #28773416 - 05/13/24 07:27 PM (1 month, 13 days ago)

Quote:

Yuggoth said:
> The jar on the left has always been kind of cloudy...

ALWAYS cloudy, like, from the time it came out of the PC?

I have made LC that was a little cloudy when freshly sterilized, due to ingredients. But if it was clear and it turned cloudy after inoculation, that is bad news.




I can’t remember for sure but I think it became cloudy when I injected the Myc. When I sterilized the broth (same for both jars), I also sterilized some broth inside a syringe in the PC. When it was done I used the Josie poke method to inoculate left jar. I want to say it became cloudy at that point but like I say can’t remember for sure.

I’m in my first series of grows so running various experiments like this. I will try inoculating grain using the jar on the right.

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Offlinevicepope
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Re: Is this LC ready for grain? [Re: Myco816]
    #28773617 - 05/13/24 11:29 PM (1 month, 13 days ago)

Test on grain if already cooked, the one on the rights looks normal. If you got agar made test it on that.  You got pf cakes made test on that. A jar of grain isn't big lost same with 1 pf cake.

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InvisibleNichrome
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Re: Is this LC ready for grain? [Re: Myco816]
    #28773676 - 05/14/24 01:15 AM (1 month, 13 days ago)

Yeah the one on the right might be good. The one on the left is turbid and not good. Don't bother even testing that one just toss it.

I like to drip some on a dish at the time of using it with grain. That way I am not only testing the LC but I am testing the whole inoculation run, the syringe, the atmosphere, etc, at the time of use. Two birds with one stone. You do run the risk of putting contaminated LC on grain and losing that grain by doing so but I like it because it reduces the number of moves to get to the end and therefore reduces the number of vectors for fucking up but still tells you what you need to know. A test plate for just the LC is groovy ahead of time but it only tells you the state of the LC at that "move" and is a good move if the LC looks or is suspect to be off.

Every time you open or toggle or use the LC in any way you could be introducing contamination. If you test LC days ahead of time it is still a good idea to do another control plate at the time of inoculation. That way you test the noc run and the lc that could have been contaminated at the time of testing it once already, even if that test plate proved clean. Each "move" or toggle is a potential vector for fucking up. Hope that madness makes sense as my words at the moment are not so becoming.

Never be afraid to test things on agar, streak or smear things on agar, deposit or drop things on agar. Agar is great, and every juncture you transfer is just dandy. Agar is the middle and the tell. 

A plate will tell you the truth of the time the plate was made but a plate can never tell you the future. Plates are echos of a moment that has passed. A moment captured. Plates tell multiple layers of truths if you learn to listen.


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Offlinevicepope
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Re: Is this LC ready for grain? [Re: Nichrome]
    #28773681 - 05/14/24 01:26 AM (1 month, 13 days ago)

100% agree. Every injections in and out is a vector. But I keep spare jars around to test all the time.

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