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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #29022697 -

Quote:
Stipe-n Cap said:
The odds of tissue revival is close to zero due to the temps reached in the dehydrator. Follow regular cloning practices.



This is my current understanding as well, I'm just fishing for something i may not have heard of yet. Thanks for the input.

Quote:
https://www.shroomery.org/forums/showflat.php/Number/28789802#28789802

Have you been there yet?



Yes I have. Probably not every post, but certainly the gist. I have successfully grown out spores from dry material.


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #29022700 -

Stipe-n Cap said:
Why the heck would he use serial dilution?
Or polysorbate? Why any of that?

@LN

The odds of tissue revival is close to zero due to the temps reached in the dehydrator. Follow regular cloning practices.

Serial dilution would be total nonsense. Also, vent your pressure cookers, folks.:facepalm3:

https://www.shroomery.org/forums/showflat.php/Number/28789802#28789802

Have you been there yet?




Ok stipe, you wanna battle? Serial Dilution reduces and separates colonies making isolation easier. Polysorbate further breaks apart clumping. You of all people should know this considering you're the one who introduced it to me. Taking samples from each stage of dilution increases your odds.

And I said his chances of success were not likely.

Vent your PC or not and tell me if you notice a difference. What's next stipe? 45 minute agar cooks lmao. Your wasting people's time.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight] * 1
    #29022702 -

Right on.

So as you saw in that post I had some rhizo on one of the germ plates, which may have indicated clone material was revived. I wish you the best in this endeavor, should it work out make sure to post over there so I can see the results. Pretty please.

@supa

I don't want to fight about flat earth or any of that other shit either. I'll leave you to your own musings.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #29022707 -

Lol no worries I'm with you on the earth being round. I'm not blowing spit bubbles yet today.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Radamanto]
    #29022709 -

Radamanto said:
I’m only asking/researching about enmeshed molds because my cultures started looking weird. My first batch of plates all looked tomentose with 0 rhizo growth but I could tell they were cubes and even grew decent pf cakes. On this new batch almost all my transfers are looking like this:



The myc used to grow on the surface of the agar but now it’s fuzzier, and growing a bit upwards, more tridimensional growth.

To be fair, I did change my agar recipe a bit, my first batch was 2% nutes 2% agar and the new is 1% nutes 1.5% agar (I was trying new mixes to get some rhizo growth) and also the new plates are cheap chinese ones so they are a bit suspicious.

@LadysKnight does this look like what you described as enmeshed mold?



It does. How far is this plate from spore? Have you made any transfers? What does the plate this one came from look like?


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers] * 2
    #29022718 -

SupaThaRipper said:
In a chance that there could be residual, dry spores, I would take a piece of exterior tissue to 2ml centrifuge filled with polysorbate water and blend on a vortexer. Let sit for 24 hours and then proceed with serial dilution to water agar.

Don't get your hopes up tho



While I don't have these fancy tools, I believe I understand their purpose in this context and I will attempt a ghetto diy version, as I have nothing to lose. Thank you


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers] * 1
    #29022719 -

It was a tongue in cheek equivalence I was attempting to make between flat earth and everything else you've been rambling about with so much self assurance.

Anyways, I've gone off topic.

Carry on, Y'all.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap] * 1
    #29022725 -

Stipe-n Cap said:
Right on.

So as you saw in that post I had some rhizo on one of the germ plates, which may have indicated clone material was revived. I wish you the best in this endeavor, should it work out make sure to post over there so I can see the results. Pretty please.




Thanks for your thoughts. I'll certainly brag on your thread if the stars align.


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

Extras: Filter Print Post Top
Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight] * 2
    #29022726 -

beginning to think I have yet to make a plate that ISN'T all enmeshed mold :lol:


--------------------
Learn it, know it, live it


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap] * 1
    #29022727 -

Don't worry, I understood. As long as you keep the words to 8 letters or less, I can usually somewhat understand.

@ladysknight, I love it. Don't ever be afraid to try anything. You don't know if it works unless you've tried it yourself. Regardless of what people say

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers]
    #29022735 -

Dumb agar question:

Should I filter out the solids after mixing up a batch of MEA? My PDA plates are totally opaque, and I just recently saw Milkboy's PDA thread, and it got me wondering if filtration would help with MEA plates. I haven't done any yet, but I need to get some going this week.


--------------------
"For the powerful freedom-takers who rob justice from us all, that's why I keep my pistol loaded, powder, cap, and ball..."
Read more, post less!
...turns out, I'm an amateur at everything

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Re: AGAR ENVY! (Anything and All things agar!) [Re: KlarnEvil9] * 2
    #29022760 -

If there's noticeable solids, I would stir until dissolved. If they don't dissolve, then yes, filter. Otherwise you need to increase your sterilization time. It takes longer to penetrate those solids. Otherwise you increase your chances of contamination

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Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight]
    #29022765 -

This is a T4 from spore. I don't have a picture of my T3 right now, but here is the T2 of this culture line.




T3 looked better, slightly more rhizo and stronger, thicker threads. Just for comparison, this is a T3 from a different  culture line, but the T3 from which I transferred the blue plate in the previous post looked almost like this but a bit better.



I’m cleaning clones of my best pf cake flushes in brf pucks right now, I also have some cakes ready to harvest soon, would you recommend to just toss the line with the enmeshed mold and start again from those other options?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhenoDreamers] * 1
    #29022769 -

Sweet, thanks for the info!
SupaThaRipper said:
If there's noticeable solids, I would stir until dissolved. If they don't dissolve, then yes, filter. Otherwise you need to increase your sterilization time. It takes longer to penetrate those solids. Otherwise you increase your chances of contamination



Sweet, thanks for the info! I hadn't considered the solids needing more sterilization time.


--------------------
"For the powerful freedom-takers who rob justice from us all, that's why I keep my pistol loaded, powder, cap, and ball..."
Read more, post less!
...turns out, I'm an amateur at everything

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Radamanto] * 1
    #29022776 -

Before you go too far worrying about any persistent contamination, I believe you may want to tighten your standard procedures. Yes, they don't look great, but at the same time, you waited too long to take those transfers. The longer you wait, the less likely you can transfer without contamination. If you followed the same path on your germination plate, you've made it harder on yourself by giving everything time to merge together. Try taking transfers as soon as you see something clean and uniform - that you can grab without also getting pieces of what you're trying to leave behind.


--------------------
It's so easy to laugh
it's so easy to hate
It takes guts to be
gentle and kind
-the smiths

Your ego is your nemesis.
Seek the truth tirelessly,
Even at the cost of your own sanity.
-fiddle_head

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Re: AGAR ENVY! (Anything and All things agar!) [Re: KlarnEvil9] * 2
    #29022793 -

KlarnEvil9 said:
Dumb agar question:

Should I filter out the solids after mixing up a batch of MEA? My PDA plates are totally opaque, and I just recently saw Milkboy's PDA thread, and it got me wondering if filtration would help with MEA plates. I haven't done any yet, but I need to get some going this week.




I use a stick blender. Go full throttle for a minute.
However, I’m using ELME, it’s pretty clear even without the stick blender. I cook it for 4 minutes before PC’ing for 25-30 minutes depending on which bottles I use.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MycCycle]
    #29022842 -

MycCycle said:
KlarnEvil9 said:
Dumb agar question:

Should I filter out the solids after mixing up a batch of MEA? My PDA plates are totally opaque, and I just recently saw Milkboy's PDA thread, and it got me wondering if filtration would help with MEA plates. I haven't done any yet, but I need to get some going this week.




I use a stick blender. Go full throttle for a minute.
However, I’m using ELME, it’s pretty clear even without the stick blender. I cook it for 4 minutes before PC’ing for 25-30 minutes depending on which bottles I use.



:fingerpistol:

That might be the ticket! I have a trusty stick blender that has blended many blendables over the past 2 decades... I don't know why I haven't thought to use it for this. Thank you!!!


--------------------
"For the powerful freedom-takers who rob justice from us all, that's why I keep my pistol loaded, powder, cap, and ball..."
Read more, post less!
...turns out, I'm an amateur at everything

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Re: AGAR ENVY! (Anything and All things agar!) [Re: KlarnEvil9]
    #29022846 -

Is this white spot bacteria or something else?
From what I've seen people say, I should transfer from the opposite side of the plate, but I'm also wondering if you see signs of bacteria mixed in anywhere with the myc?
Does the contam only exist in the one spot or is a spot like that just one visible part of a much bigger and less visible contamination?



Also for this yellow spot. Bacteria right? And the same question as above, does it likely just exist in that one spot and would be spreading from there, or likely in more places and only visible in that one spot?

Edited by Butter_Butter (11/13/24 01:02 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Butter_Butter] * 1
    #29022855 -

yup bacteria. you should be able to transfer from the opposite side of it, i wouldnt but the rest of the plate on grains, just transfer


--------------------
I call them Huskies cause they Mush!


The Official Wild Cubensis Thread
Instant Pot Agar
Gnocchi Water Agar
Let's Talk Cowtek

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Mackload]
    #29022879 -

Mackload said:
beginning to think I have yet to make a plate that ISN'T all enmeshed mold :lol:



An interesting exercise is TRY to grow some mold and bacteria.

Open an agar plate for about 30 minutes in the room where you work. Not in a SAB, just in the room. Do not add spores.

Seal the plate. Open another plate, smear some dust across it.  Do not add spores. Seal it up.

Don't open the plates again, let them grow out, keep sealed, photograph, then toss.

Watching what grows might help recognize what is contam from your environment and what is not.

One of my first plates, summer 2020.  I wasn't trying to grow this, but later I tried and it looked about like this. White is shroom mycelium. Orange is bacteria (there is no food coloring in this plate.) Green is mold. Below that is a good plate from 2023.

That is one of the worst plates I've seen that also did have shroom mycelium.

Good plate:

Edited by nektar61 (11/13/24 01:50 PM)

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