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The Tentative Crucible Tek - Running Spores through the Pressure Cooker * 4
    #29691804 -

This is stupid, but I have evidence, so hold your horses and open your mind, maaan.

There was a thread about cleaning spores using the dehydrator. Didn't pan out well for anyone from what I remember.

So I decided to do something stupid, because obviously the PC kills everything, right?

Well, as we know, that is not always the case. Spores should not have survived, but between being in a thick plastic pp5 container, it being an Instant Pot that doesn't get to 15psi, getting smeared at the bottom of the container which had an air gap above the water, me using an entire spore print, and using the Yeti swab to swab everything, and then smear it at the bottom of the container, and probably a safe area due to the protective barrier of their siblings cellular corpses, something survived.

Note: if you are using a regular autoclave, maybe stop it at 11psi or so.

Steps Taken for First Tentative Crucible Tek:
1- Used a swab with Yeti to swab up an entire Husky sporeprint.
2- Rubbed the mixed up swab against the bottom of my pp5 plastic container, 100ml, snack container, exactly this model
3- Wrapped it in foil, and perhaps used a protective paper towel layer on top a la pastyplates, don't remember.
4- PC'd with Agar for 45 minutes in the 8 quart Instant Pot
5- Waited for a long time.
6- Noticed growth in the form of a mycelial conga line surrounded by visible spore ornaments

Both Yeti and Husky phenos showed up, perhaps not intertwined but separate, not sure. Gonna see what my two last shoeboxes pop out.

Due to the ruptured cells of the majority of the spores, that water became a low nutrient solution, perhaps enough to induce germination. Maybe it was just the water itself.

The jar was sitting for weeks/months, and right when I was about to dump it, I saw a mycelial colony. I plopped it on agar in open air.



It contaminated where it landed, but i put it to the side, and it grew well



It overcame the contamination, which I think was a yeast. I think it was in the original container but not sure.



For crosses, heterosis, aka hybrid vigor can be a thing and I think this was an example of it, although it fruited terribly, perhaps because it was never truly clean and had enmeshed nasties from the get go.



Gotta admit, there were a bunch of questions for AI for in depth niche mycology, but what I gathered, is that it might have been a way for selecting spores with strong hydrophobin genes.

Hydrophobins can protect against heat, help create little air pockets where maybe spores could survive. They are what cause aerial mycelium, letting it move upwards. They are also what causes droplets on the surface mycelium, since they stop water from entering the cell. Actually, they are the gatekeeper, allow in or out, from what I remember.

Yusky mycelium went VERY TALL on agar plates. Effectively skyscrapers at scale, indicating to me that the Crucible Tek is a way for selecting for that.





So basically, the IP with the plastic armor spaceship created an environment where probably something like 99.99% of spores were annihilated, but under their corpses, some survivors managed to survive.

Unfortunately, it was either too contaminated, or it truly fruits like shit. Never got a proper flush, although did get a few micro fruits. I did a island isolation on agar to try to clean it up and that seems to have helped. Got two shoeboxes of it atm and hopefully those pan out.

I tried again, didn't get good results. A lot of the containers I used warped a little, or had a hole on top from pasty plate, and perhaps just a thin micropore allowed contaminants in.

Latest Experiment:

I used Eungella with Purple Mystic, put it in a glass heinz bottle with a few drops of maple syrup, and PC'd it for 20 minutes in the Instant Pot, without venting.



Physics made most the spores go to the circumferance of the bottom, which is great, since maybe it is there, covered in mounds of other spores, that certain freaks can survive. I'm going to leave it not agitated for a while, and then later try shaking it around, and see if stuff starts growing. Not sure how I will remove it from the bottle but first lets see if anything happens.

What I want to discover, is if we can find the sweetspot where enough spores reliably survive without bacteria or molds doing the same. This would allow for sterile LC, including F1 crosses.

Anyone down to PC some spores with me?


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


Huskies' Grimoire
The Official Wild Cubensis Thread
Easy Instant Pot Agar
Let's Talk Cowtek

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies] * 1
    #29691819 -

Quote:
Anyone down to PC some spores with me?



Hmmmm okay. my neglected tidal wave is absolutely puking spores, and the instant pot isn't busy. Fuggit!


--------------------


.~`  ๐”… ๐”ž ๐”ค ๐”ข ๐”ฉ  ๐Ÿฅฏ  ๐”… ๐”ฆ ๐”ฑ ๐”ข ๐”ฐ  `~.
:salute: CAKE WARS III HAPPENING NOW :salute:

๐Ÿงช Castellani Preservation | ๐Ÿ”ฌ The Slidewich | โ˜ ๏ธ Touchless skull sterilizer | ๐Ÿ“ธ Agar plate scanner


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Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Rasmodius] * 1
    #29691833 -

Honestly, you raise an interesting point. Even if it's as deliberate stress applied to spores for selection purposes.

It reminds me of using sublethal UV to force mutations โ€” I don't know if anyone around here has tried it โ€” though it wouldn't surprise me.

Maybe I could test it, but as you know I don't have a PC, but... You could prepare several spore suspensions in distilled water and steam them for different lengths of time, or straight-up cook them in a water bath at various times and temperatures.

In fact, I find that way more interesting than jumping headfirst into temperatures >100 degrees Celsius. At 40, 50, or 60 degrees, things must happen too.

And what about freezing in an aqueous suspension?

Hugs :heart:


--------------------
"Little shark in da bathtub"


Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Rasmodius]
    #29691866 -

Rasmodius said:
Quote:
Anyone down to PC some spores with me?



Hmmmm okay. my neglected tidal wave is absolutely puking spores, and the instant pot isn't busy. Fuggit!


:punker: what container are you thinking? half pint jars and pp5 containers seems like the best options.

I do think that swabbing it on the bottom can be beneficial, but I did not this last attempt.

@belintaf

That is a very good point. Simply boiling could already go most of the way, but I would say that would only be for cleaning up syringes for MS.

That's an easy enough experiment though

1- Get a non ideal dirty print, or even leave one in open air
2- Put it in a pint container of water.
3- Grab one syringe for testing base cleanliness either on agar but PF Tek would be better.
4- Separate into smaller containers or even use individual syringes and boil those directly.
5- Take one out at  the 10, 15, 20, 30, 45 , 60 minute intervals
6- Inject onto plates or cakes, see if you still get viable spores that grow, and which timing did mold and bacteria issues not appear to matter for getting a clean cake.

Thing is, we know that without sterilizing, bacteria endospores and mold are more resilient. Dont want to grow 12 mold jars on purpose.

It might've been the unusually large amount of spore mass that turned the tables.


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


Huskies' Grimoire
The Official Wild Cubensis Thread
Easy Instant Pot Agar
Let's Talk Cowtek

Edited by Huskies (10/09/26 04:31 PM)

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies]
    #29691880 -

Hahaha, when I was reading your post, those exact things were crossing my mind. :lol:

And yeah, we definitely need to test it out. I have a huge Golden Teacher print I made to try making ink, which has been sitting open on the desk for a few days. It could be a great candidate.

The problem is that right now I don't have any jars, small pots, or compact containers left. But next week I'll go buy some.

I love the idea! :evil2:


--------------------
"Little shark in da bathtub"


Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Belintaf]
    #29691883 -

Great! How'd the ink turn out?


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


Huskies' Grimoire
The Official Wild Cubensis Thread
Easy Instant Pot Agar
Let's Talk Cowtek

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies]
    #29691890 -

There's the print waiting โ€” but you know what? You've got me hooked.

I'm going to do it right now. I was going to do it next week because I need to buy Tween 20 to use as a surfactant, but... whatever. I'll redo it with Tween 20 later on, for now I'll use dish soap diluted to 0.1%. And agar as a binder at ~0.3%.

I'm on it โ€” I'll check back in a bit. :lol:


--------------------
"Little shark in da bathtub"


Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Belintaf]
    #29691899 -

Selecting for hydrophobin spores might bring along with it a whole host of other extreme environmental genetic defences

Maybe the hydrophobin spores also have a dormant/innate ability to fight off extreme cold temperatures like Belintaf suggests...

I am going to do these tests with the new spores I am getting :thumbup:

I shall report back with my findings

:salute:


--------------------
"Why try for high fives,
When my pies have nine lives?"
DUBJUG : H.D. : PLASTIC-MYC : TGE : JOIN CAKE WARS : MODERN FRUITING : MCWA : QDMT : โ™พ๏ธ : MY FAV TEKS ๐Ÿ“–

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies]
    #29692015 -

Huskies said:
:punker: what container are you thinking? half pint jars and pp5 containers seems like the best options.

I do think that swabbing it on the bottom can be beneficial, but I did not this last attempt.


I have some small glass jars with plastic screw lids that have survived multi-hour PC cooks. I'm not sure if they're PP, got them from a secondhand store, they're the type of thing small creams or cosmetics would come in. My PP condiment cups have been deforming lately. They still seal, but it's enough that I'm shying away from them on anything LC related bc I don't wanna deal with the leak/contam risk.


--------------------


.~`  ๐”… ๐”ž ๐”ค ๐”ข ๐”ฉ  ๐Ÿฅฏ  ๐”… ๐”ฆ ๐”ฑ ๐”ข ๐”ฐ  `~.
:salute: CAKE WARS III HAPPENING NOW :salute:

๐Ÿงช Castellani Preservation | ๐Ÿ”ฌ The Slidewich | โ˜ ๏ธ Touchless skull sterilizer | ๐Ÿ“ธ Agar plate scanner


Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Rasmodius] * 2
    #29692061 -

Haha, well. Getting back to the ink topic... I'm starting to think this could actually be viable.

I made it like I said. 0.3g of agar (binder) in 100ml of water with 0.1% dish soap (surfactant) as a base, plus that dense spore print.



I scraped the entire print into the shot glass and added the solution using a syringe โ€” I went a little overboard with the amount. If I had added less, the ink would have turned out really dark, but it didn't turn out bad at all.



I don't have a pen, but I'll make one. I improvised with whatever I had on hand. Though even just a couple of lines on a card could be enough.

I doubt spore viability will hold up when using dish soap, but it costs nothing to check. With Tween 20, that issue should disappear.

Damn โ€” it looks really good. :awesomenod:

Hugs :heart:

Edit:

Damn, once it dries โ€” it holds up really well. When trying to scrape it with the tip of a knife, this was the result:





Edited by Belintaf (10/09/26 06:56 PM)

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Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Belintaf] * 1
    #29692683 -

:sexymeow::havesomescience:


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


Huskies' Grimoire
The Official Wild Cubensis Thread
Easy Instant Pot Agar
Let's Talk Cowtek

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies] * 1
    #29696642 -

One of the last ones I thought had failed seems to have created multiple viable clumps, to the naked eye I saw mycelium for sure



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


Huskies' Grimoire
The Official Wild Cubensis Thread
Easy Instant Pot Agar
Let's Talk Cowtek

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies]
    #29697925 -

Went pearl diving.



Really easy to pick them with sterilized tweezer, they ball into a droplet (hydrophobin forcefield?) and then lay to rest easy on agar.

This is Yeti x Murbah Australia, so may I introduce you to the tentative cross, Dropbear



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


Huskies' Grimoire
The Official Wild Cubensis Thread
Easy Instant Pot Agar
Let's Talk Cowtek

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Huskies] * 2
    #29698476 -

Huskies said:
There was a thread about cleaning spores using the dehydrator. Didn't pan out well for anyone from what I remember.


Yes, that was my thread. It works quite well for me, hobbit, pheno, and likely others. You kind of have to know what you're doing; I'd call it an advanced technique, it's not fool proof but remains an effective tool nonetheless.

Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Stipe-n Cap]
    #29698573 -

Thanks Stipe-n-Cap. :japsmile:

I didn't know about your thread โ€” if I'm not mistaken, it's this one โ€” Spore cleaning without agar or centrifuge (pasteurized spores).

I just read through the whole thing, but I couldn't find a clear conclusion.

Would you mind sharing what you guys concluded about it? Or at least share a link where I could read more on it? I'd really appreciate it, to be honest.

And it definitely seems like a super interesting topic.

Hugs :heart:


--------------------
"Little shark in da bathtub"


Extras: Filter Print Post Top
Re: The Tentative Crucible Tek - Running Spores through the Pressure Cooker [Re: Belintaf] * 2
    #29700835 -

I won't derail this thread other than to post SH's reply to that thread re his success. I'll be updating the OP to reflect current procedures when I can settle into a place to work.

https://www.shroomery.org/forums/showflat.php/Number/29195082#29195082

Didn't mean to derail your thread, Husky. Any further questions should be asked in the other thread.

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