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Modern day mycelial slurry syringes


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#1 cybele

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Posted 17 September 2019 - 04:05 PM

I am in the process of fabricating some plastic lids.

 

The plan is to have plenty of clean bulk inoculate using the Mycelial Syringe tek. After I isolate a good culture on agar I would like to then transfer a clean wedge to a sterile grain jar. The grain jar will then be used to create mycelial syringes. I plan to follow the tek to a T with the exception of making the SHIP obsolete. I plan on  doing that by making some changes to the lids.

 

I am using the following:

  • half-pint jar
  • wide-mouth plastic lid, grey
  • self-healing injection port, translucent/clear
  • syringe filter .22 micron pore size

The syringe filter should stop the jar from coming under vacuum. If I feel resistance when pulling the sterile water back up then I know we need more air flow. If that occurs I can place a ship through the same injection port as they are built for longevity. Either way we are all going to find out!

 

81yv6p-Mwy-JL-AC-UY218.jpg

[This is the exact design, my parts are just different colors. I will post a picture of my lid's as soon as they are done, and cured.]

 

I am using the following thread's:

 

Fahster https://mycotopia.ne...ge-tek-revised/

BuckarooBanzai's mycelial slurry tek https://mycotopia.ne...-inoculant-tek/

 

 

My only question is the grain prep. I would like to use popcorn, but I have only worked with oats. Fashster went into some detail with his grain prep, but I believe his  grain hydration revolves around his tyvex absorbent lids. What is the best way to prep, and hydrate popcorn, on an extremely small scale. I will still be using oats as my primary spawn. I am just using the popcorn to "shred" the myc for the syringes.


Edited by cybele, 17 September 2019 - 04:06 PM.

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#2 sandman

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Posted 18 September 2019 - 07:53 AM

Don't forget a silicone gasket for the lids. Plastic lids are typically not water tight because they have no gasket area. Use the thick clear FLAT silicone gaskets and NOT the Tattler brand.  


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#3 cybele

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Posted 18 September 2019 - 10:49 PM

Don't forget a silicone gasket for the lids. Plastic lids are typically not water tight because they have no gasket area. Use the thick clear FLAT silicone gaskets and NOT the Tattler brand.


Thanks for the advice sandman. The inner design is different on my newer grey lids. They are the ball brand, and differ from the white lids I have as well that are pictured above. However, I will still be picking up some gaskets so thanks for saving me the future headache Any input on the popcorn prep? .

#4 DetritusTheEgo

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Posted 19 September 2019 - 12:19 AM

Pretty sure that if you run with regular mouth plastic lids vs wide mouth plastic lids you can get away without a gasket with that lid configuration. Shroomsupply sells both regular mouth aka 70mm and wide mouth aka 90mm lids in those configurations and they state to use a gasket with the 90mm and not the 70mm if your going after a watertight seal. I use those lids in wide mouths with a gasket, but just a heads up.

This is pretty much inline with what I do with whole corn: https://mycotopia.net/topic/63118-turkeyranch%E2%80%99s-easy-popcorn-spawn-pictorial/

As outlined in that tek, if you're worried about getting the right hydration you can add a few tbsp of vermiculite to the bottom of each jar to soak up small amounts of excess water before pressure cooking them. Use that as a "bandaid" until you dial in your process if you have to.


Edited by DetritusTheEgo, 19 September 2019 - 12:24 AM.


#5 cybele

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Posted 19 September 2019 - 07:28 AM

This is pretty much inline with what I do with whole corn: https://mycotopia.net/topic/63118-turkeyranch’s-easy-popcorn-spawn-pictorial/
As outlined in that tek, if you're worried about getting the right hydration you can add a few tbsp of vermiculite to the bottom of each jar to soak up small amounts of excess water before pressure cooking them. Use that as a "bandaid" until you dial in your process if you have to.

Thank you Detritus!! That thread is exactly what I needed. I cannot use the vermiculite bandaid as it will interfere with the syringe drawing up the sterile water with mycelium. I am just going to have to perfect the hydration, and that tek made it seem real simple.

Edited by cybele, 19 September 2019 - 07:30 AM.


#6 FunG

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Posted 20 September 2019 - 03:14 PM

I use popcorn, the brand I get is "no name" it's easy, just fill the jars half way, add water up to the rim of the jar, wait 12-24 hours and it's at field capacity.

Pressure cook and upon cooling there is always pooled water at the bottom of the jar but just tilting the jar upside down and rolling the popcorn helps distribute the water back into the kernels. It gets reabsorbed....no need to add vermiculite.

The first time I used popcorn the pooling water threw me off and caused a panick, thankfully I remembered the hot/cold expansion rule.

Your lids are awesome, I seen that design posted on another forum before. I want to make some but dont know what to use them for since I g2g everything. And as for the mycelium slurry, I attempted it once before using sterilized water but for some reason I got a bacteria contaminate....this is going back 8-9years so I was still pretty new but it was enough to opt for more sterile inoculation methods.

Hope it works out for you man
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#7 cybele

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Posted 21 September 2019 - 08:58 AM

I use popcorn, the brand I get is "no name" it's easy, just fill the jars half way, add water up to the rim of the jar, wait 12-24 hours and it's at field capacity.

Pressure cook and upon cooling there is always pooled water at the bottom of the jar but just tilting the jar upside down and rolling the popcorn helps distribute the water back into the kernels. It gets reabsorbed....no need to add vermiculite.

The first time I used popcorn the pooling water threw me off and caused a panick, thankfully I remembered the hot/cold expansion rule.

Your lids are awesome, I seen that design posted on another forum before. I want to make some but dont know what to use them for since I g2g everything. And as for the mycelium slurry, I attempted it once before using sterilized water but for some reason I got a bacteria contaminate....this is going back 8-9years so I was still pretty new but it was enough to opt for more sterile inoculation methods.

Hope it works out for you man


I appreciate the advice with the popcorn. It could not sound easier! I am sure I will learn with some trial and error! The lids are coming together, unfortunately they sent me the wrong size syringe filters. I hate that yours got contaminated, but at least you could spot it unlike in Liquid culture. Hopefully these lids will narrow my chances of contamination.

The idea is to save me from making constant agar transfers to other agar dishes, and to grain. Once I isolate a culture I should only have to transfer a small wedge to the mycelia slurry jar. Then the syringes I make should inoculate the first tier of jars. Then I will G2G from those. Making my mycelia popcorn jar the “master” Instead of chopping up the entire plate and doing grain transfers. Or inoculating one master grain jar with one wedge, and then waiting to G2G. I still have some figuring out to do with the processes but that’s the jist.

Edited by cybele, 21 September 2019 - 09:12 AM.

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#8 cybele

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Posted 05 October 2019 - 08:38 AM

The new syringe filters came in. Time to fabricate these lids, and let them cure.

After curing, we are going to whip up some popcorn. Then it’s time to put a little agar love in the jars. After colonization we will find out if the syringe filter is adequate enough, and there is no need for a ship.

I’ll be transferring clone tissue to agar, then to popcorn, then the mycelial syringe to grain with my cubes. I also plan on transferring pan agar to the popcorn master, and using the mycelial syringes to perform waylitjim’s cyan tek.


41285282-8-DA5-491-E-9-B4-C-054-C0-DAA94

Edited by cybele, 05 October 2019 - 08:39 AM.

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#9 cybele

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Posted 05 November 2019 - 01:28 PM

All of the lids came in, and I fabricated everything up. I have grain lids, and lids for the popcorn tek all cured. The time is closing in for some possible Golden Teacher clones, and after agar isolation I plan to transfer to a popcorn jar for bulk inoculate.

 

With the time closing in to really use these lids, I want to test them. So this weekend I am going to hydrate and sterilize some popcorn in multiple jars. After, I will transfer some cultures to them. I have some less than worthy Cyan cultures that will fit the bill for test subjects. This will give me the chance to test the air diffusion with the syringe filters, and see how contaminant tight these lids are before I waste any cultures.

 

It is going to get fun from here.



#10 FunG

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Posted 06 November 2019 - 01:48 AM

I'm not to sure how pan cyan will do on popcorn.... popcorn is not the greatest grain as far as nutrition goes, cyans may not preform well on popcorn.id recommend rye for the cyans, that's the most common grain people grow them on.

Popcorn works "o.k" with p.cubensis but the nutritional value is nowhere near that of rye or wbs, just dont want to see you wasting your time.
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#11 cybele

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Posted 07 November 2019 - 07:26 PM

I'm not to sure how pan cyan will do on popcorn.... popcorn is not the greatest grain as far as nutrition goes, cyans may not preform well on popcorn.id recommend rye for the cyans, that's the most common grain people grow them on.

Popcorn works "o.k" with p.cubensis but the nutritional value is nowhere near that of rye or wbs, just dont want to see you wasting your time.

 

Right on FunG. I love how clever your name is of a double entendre!

 

Thanks for the advice! I am from a grain state, and we have access to many types here. Which is another reason I beat myself up for not moving to bulk sooner, but my career used to have an odd travel schedule. I do try and stay versed in their nutritional values however. To clarify I am not growing my cube or cyan species "on" popcorn. I will be using oats with some gypsum, and a small amount of popcorn powder as my spawn.

 

I will be utilizing the popcorn to produce a bulk inoculate. In place of transferring an agar wedge into one grain jar, and then transferring that to other grain jars. Instead I will take an agar jar with an isolated culture, and i'll transfer a wedge into the popcorn jar. I will then add some sterile water, shake vigorously, and draw up the shredded mycelium liquid into syringes that will be used to inoculate multiple jars at the same time. From my understanding popcorn was found to shred the mycelium up best. 

 

I am sure you can use many different grains, place some marbles, glass, or any other improvised shard device, and it will work. Like everything in this hobby there are many different ways to do the same thing. With me easing into working with bulk I am still setting up my procedures, and some things are bound to change along the way!



#12 cybele

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Posted 14 November 2019 - 07:42 PM

Alright, alright, alright. All three cultures are showing signs of a successful transfer! Mycelium is spreading and there are no signs of contaminants so far. More to follow!

 

IMG-1485.jpg

[48 hours from transfer]


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#13 flashingrooster

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Posted 14 November 2019 - 08:58 PM

Was it silicone that you were curing?


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#14 cybele

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Posted 14 November 2019 - 09:19 PM

Was it silicone that you were curing?



Yeppers! I use permatex clear silicone sealant. I come from an aviation background, and sealant is used every single day for a plethora of uses. I stay away from the high temp as the clear is more malleable and can withstand 400 degrees Fahrenheit sustained.

One easy tip I have is to keep windex close by. The alcohol percentage is not enough to degrade the silicon and it prevents the silicone from sticking to your finger, gloved or not, or whatever device you use to apply your sealant. If sealant gets somewhere you don’t want it to be you can use a paper towel/rag with windex and wipe it right up without spreading it.

24 hours is the known cure time for most clear sealants. I let mine go for 48 just because of humidity variables etc. I don’t let anything I have recently sealed sit until use. After the 48 hours it goes through a heat cycle that I believe helps mold the silicone better to shape, and sets it in place pushing out air bubbles that are hard to see.

The silicone will change through out seven days even if it only takes one to chemically “cure”. You can feel this by touch after working with enough of it.

No silicone is visible in my construction, the minimal amount is used, and that prevents degradation in the future.

#15 cybele

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Posted 14 November 2019 - 09:24 PM

Don't forget a silicone gasket for the lids. Plastic lids are typically not water tight because they have no gasket area. Use the thick clear FLAT silicone gaskets and NOT the Tattler brand.

I quoted this before Sandman letting you know I visually saw a design/engineering difference between the grey and white lids. Well here’s an update. I PC’ed 12 lids through multiple size jars, and didn’t get any water in or out.

I tightened the lid up snug, then backed it off, essentially trying to do as little as possible to break the seal. The lid’s were also not wrapped with foil.

Edited by cybele, 14 November 2019 - 09:25 PM.


#16 FunG

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Posted 15 November 2019 - 02:31 AM

Thank you cybele you made my night, I never thought anyone would take notice to the handle.

Its suppose to be a cute way my friends kid says fung"I" but prounces it "fungee"

I understand what you're doing now too, thanks for explaining yourself, popcorns surface space is larger so more mycelium would run off when introduced with water.

Still, slurry syringes make me sketchy just from the experiences I've had with them but they were attempted from rye grain jars prior to me even knowing about g2g.
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#17 macgyver

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Posted 15 November 2019 - 03:44 AM

I did a popcorn jar not too long ago to make slurry! Did a pint and inoculated with LC, It took 12 days to colonize fully. I could definitely have used these lids... I just had a hole stuffed with poly, removed the poly and shot 120ccs of sterile water, shook like hell and extracted. I've used it to inoculate a batch of BRF and a batch of grain jars and no problems as of yet.

 

I'll definitely be revisiting the popcorn jar once I have a nice wedge to use. I'll probably have to revisit this thread and steal your lid design too :biggrin:


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#18 Sidestreet

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Posted 15 November 2019 - 04:51 AM

 

One easy tip I have is to keep windex close by. The alcohol percentage is not enough to degrade the silicon and it prevents the silicone from sticking to your finger, gloved or not, or whatever device you use to apply your sealant. If sealant gets somewhere you don’t want it to be you can use a paper towel/rag with windex and wipe it right up without spreading it.

 

Cool, I need to try that.  Maybe next time I use silicone I won't end up with a sticky mess.


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#19 cybele

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Posted 15 November 2019 - 03:54 PM

I did a popcorn jar not too long ago to make slurry! Did a pint and inoculated with LC, It took 12 days to colonize fully. I could definitely have used these lids... I just had a hole stuffed with poly, removed the poly and shot 120ccs of sterile water, shook like hell and extracted. I've used it to inoculate a batch of BRF and a batch of grain jars and no problems as of yet.

 

I'll definitely be revisiting the popcorn jar once I have a nice wedge to use. I'll probably have to revisit this thread and steal your lid design too :biggrin:

 

Congratulations on your success thus far! That is what I love hearing!

 

There are countless methods to every part of this madness, and for myself it is what makes this hobby so interesting. I am fixing to start a unique travel schedule with work. Being able to have everything started at the same time is the best route for me. Compared to staggering it out with grain to grain transfers.

 

I did not design the lid, but thank you on the handy-work. I can only assume that the design started before the internet, and easy access to these affordably manufactured medical products.

 

The old design was two holes drilled in a lid with silicone LIBERALLY added to seal the hole on both sides of the lid. One silicone hole was used as the self-healing injection port for your syringes.The other was used to place a syringe used as a "ship" to displace pressure. The design was ingenious, and to this day requires absolutely no improving. I kept the same engineering principles, but added parts for the longevity. I do not want to have to be repairing silicone when I am home, and have the time to be in the lab.



#20 cybele

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Posted 15 November 2019 - 03:55 PM

 

 

One easy tip I have is to keep windex close by. The alcohol percentage is not enough to degrade the silicon and it prevents the silicone from sticking to your finger, gloved or not, or whatever device you use to apply your sealant. If sealant gets somewhere you don’t want it to be you can use a paper towel/rag with windex and wipe it right up without spreading it.

 

Cool, I need to try that.  Maybe next time I use silicone I won't end up with a sticky mess.

 

 

Sealant used to give me nightmares before I learned that trick :ohmy:


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