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Invisiblesandman420
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How are commercial PE syringes made? * 1
    #25113828 - 04/04/18 02:11 PM (5 years, 9 months ago)

Does anyone have any info on how commercial syringes for strains that don't print are made, like PE and APE?


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InvisibleJHOVA
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Re: How are commercial PE syringes made? [Re: sandman420]
    #25113909 - 04/04/18 03:03 PM (5 years, 9 months ago)

put a sterilized swab into a sterilized jar with a ship and ge port. Aspirate that. Wanna be fancy add a drop of glycerin.
I think munchauzen was gonna make a video on making syringes if i recall correctly


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InvisibleGerms
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Re: How are commercial PE syringes made? [Re: JHOVA] * 2
    #25113921 - 04/04/18 03:10 PM (5 years, 9 months ago)

With a centrifuge is how Sporeworks does it I believe


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InvisibleLemurLemur
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Re: How are commercial PE syringes made? [Re: JHOVA]
    #25114006 - 04/04/18 03:51 PM (5 years, 9 months ago)

I wouldnt use a lid with a ge port since your gonna want to shake tf out of it.


Id use a long needle to suck em up after they settle, so use like a haft pint so it can reach.


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InvisiblebodhisattaMDiscordReddit
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Re: How are commercial PE syringes made? [Re: sandman420] * 3
    #25114466 - 04/04/18 07:32 PM (5 years, 9 months ago)

Quote:

Workman said:
It isn't really a secret but the basic method is to take very mature caps that are visibly producing spores.  Even these rarely drop decent prints but the spores are there.  These are gently dried, which tends to cause even more spores to mature before the cap completely dries out.  The caps are then crushed, rehydrated in sterile water with a wetting agent and then centrifuged.  Spores are very dense compared to the mushroom tissue, bacteria and other impurities, so they separate nicely into a pellet at the bottom of the vial.  Remove the top layer and repeat with a few more washings and centrifuging and you have nearly pure spores when you are done.  This is dried and used as needed.

It is somewhat complicated and not practical for the home experimenter but it does scale up well for the production of commercial quantities of PE spores.  This method can also be used with regular cubensis caps after normal printing.  There are a surprisingly large amount of spores clinging to gills.  Swabbing is easier and works just as well for archiving spore samples.




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Invisibleelasticaltiger
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Re: How are commercial PE syringes made? [Re: bodhisatta] * 1
    #25114528 - 04/04/18 08:04 PM (5 years, 9 months ago)

Quote:

bodhisatta said:
Quote:

Workman said:
It isn't really a secret but the basic method is to take very mature caps that are visibly producing spores.  Even these rarely drop decent prints but the spores are there.  These are gently dried, which tends to cause even more spores to mature before the cap completely dries out.  The caps are then crushed, rehydrated in sterile water with a wetting agent and then centrifuged.  Spores are very dense compared to the mushroom tissue, bacteria and other impurities, so they separate nicely into a pellet at the bottom of the vial.  Remove the top layer and repeat with a few more washings and centrifuging and you have nearly pure spores when you are done.  This is dried and used as needed.

It is somewhat complicated and not practical for the home experimenter but it does scale up well for the production of commercial quantities of PE spores.  This method can also be used with regular cubensis caps after normal printing.  There are a surprisingly large amount of spores clinging to gills.  Swabbing is easier and works just as well for archiving spore samples.







Thankyou for digging this up, I had not read that before. I always assumed they came from the occaisional "normal" looking fruits that pe sometimes puts out.


--------------------
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OfflineMr.Kaption
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Re: How are commercial PE syringes made? [Re: bodhisatta]
    #25114537 - 04/04/18 08:08 PM (5 years, 9 months ago)

Quote:

bodhisatta said:
Quote:

Workman said:
It isn't really a secret but the basic method is to take very mature caps that are visibly producing spores.  Even these rarely drop decent prints but the spores are there.  These are gently dried, which tends to cause even more spores to mature before the cap completely dries out.  The caps are then crushed, rehydrated in sterile water with a wetting agent and then centrifuged.  Spores are very dense compared to the mushroom tissue, bacteria and other impurities, so they separate nicely into a pellet at the bottom of the vial.  Remove the top layer and repeat with a few more washings and centrifuging and you have nearly pure spores when you are done.  This is dried and used as needed.

It is somewhat complicated and not practical for the home experimenter but it does scale up well for the production of commercial quantities of PE spores.  This method can also be used with regular cubensis caps after normal printing.  There are a surprisingly large amount of spores clinging to gills.  Swabbing is easier and works just as well for archiving spore samples.







:takingnotes:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: Mr.Kaption]
    #25114615 - 04/04/18 08:43 PM (5 years, 9 months ago)

for the DIY people with a 3d printer like myself i found this to be pretty interesting and might have to give it a go.

http://designawards.core77.com/Open-Design/62397/3D-Printed-Centrifuge-for-International-Health-Labs


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OfflineAsuraS
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Re: How are commercial PE syringes made? [Re: john_conner] * 1
    #25114621 - 04/04/18 08:47 PM (5 years, 9 months ago)

Quote:

john_conner said:
for the DIY people with a 3d printer like myself i found this to be pretty interesting and might have to give it a go.

http://designawards.core77.com/Open-Design/62397/3D-Printed-Centrifuge-for-International-Health-Labs




Salad spinner centrifuge :lol: Pretty damn clever.


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Invisiblesandman420
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Re: How are commercial PE syringes made? [Re: Asura]
    #25114627 - 04/04/18 08:52 PM (5 years, 9 months ago)

OK that is interesting about the centrifuge. So they take totally dried caps and crush them into some water and put that mixture into a centrifuge tube? What is the wetting agent, jetdry? I wonder how much water to crushed caps goes in the centrifuge tubes.


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InvisibleLemurLemur
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Re: How are commercial PE syringes made? [Re: bodhisatta]
    #25114640 - 04/04/18 09:03 PM (5 years, 9 months ago)

Hu! How clean can it get spore solution??


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InvisiblebodhisattaMDiscordReddit
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Re: How are commercial PE syringes made? [Re: sandman420] * 1
    #25114646 - 04/04/18 09:08 PM (5 years, 9 months ago)

could be anything there's lots of wetting agents, you can get anything you desire.
dish soap is one example of one. so is jet dry.

doesn't matter how much water. it's getting removed later anyway, use enough or more.

theoretically the PE spores should be cleanest lol


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Invisiblesandman420
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Re: How are commercial PE syringes made? [Re: bodhisatta]
    #25115244 - 04/05/18 07:05 AM (5 years, 9 months ago)

I'm guessing that the crushed caps and water mixture can't be much more than just "moistened" or it wouldn't stay separated very well after the centrifuge.


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Re: How are commercial PE syringes made? [Re: sandman420]
    #25115271 - 04/05/18 07:15 AM (5 years, 9 months ago)

Ever used a centrifuge? The compaction keeps the layers separate


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Invisibleelasticaltiger
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Re: How are commercial PE syringes made? [Re: bodhisatta] * 1
    #25115329 - 04/05/18 07:57 AM (5 years, 9 months ago)

Yeah nothing stays together in a centrefuge.


--------------------
First time growing cakes? DON'T make a Shotgun Fruiting Chamber

The Shmuvbox. - The Old TC's Like it :shrug:

Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger!
No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit.

"The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT

Pastywhite's Easy Agar Tek (PastyPlates)

Tiger Drop Video Demos By munchauzen

Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson

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Invisiblefilthyknees
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Re: How are commercial PE syringes made? [Re: elasticaltiger]
    #25115389 - 04/05/18 08:40 AM (5 years, 9 months ago)

Didn't someone once post using sterile water syringes to squirt the spores strait from the fresh cap into a container?

I heard centrifuge as well but do wonder if they used dried caps or if that is too much material.


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Invisiblesandman420
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Re: How are commercial PE syringes made? [Re: filthyknees]
    #25115400 - 04/05/18 08:45 AM (5 years, 9 months ago)

No, never used a centrifuge but I see there are some really cheap ones available. Do you think the little $50 4000rpm units would do the job?


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OfflineFishLevelMidnight
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Re: How are commercial PE syringes made? [Re: john_conner]
    #25115429 - 04/05/18 09:02 AM (5 years, 9 months ago)

Quote:

john_conner said:
for the DIY people with a 3d printer like myself i found this to be pretty interesting and might have to give it a go.

http://designawards.core77.com/Open-Design/62397/3D-Printed-Centrifuge-for-International-Health-Labs




For those of us without a 3-D printer...



How do y'all think the spores are dried? Just remove the supernatant and the non-spore bits and leave in the centrifuge tube to dry under a flowhood? Similar to an RNA pellet?

I'd like to try this with my next rounds of prints.
Any info on size and Gs needed from workman bod?

I would imagine spinning in 50 mL falcon tubes would be more efficient (It would be a PITA to add the crushed caps to 1.5 mL tubes). but I know some centrifuge inserts have a max rpm of like 4200...

Curious how easy the washing/removing the cap material is from the spore pellet. Also I wonder spore number per uL after centrifugation (or by weight of the pellet...) Sorry rambling now!


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: bodhisatta]
    #25146195 - 04/17/18 05:51 PM (5 years, 9 months ago)

Quote:

bodhisatta said:
Quote:

Workman said:
It isn't really a secret but the basic method is to take very mature caps that are visibly producing spores.  Even these rarely drop decent prints but the spores are there.  These are gently dried, which tends to cause even more spores to mature before the cap completely dries out.  The caps are then crushed, rehydrated in sterile water with a wetting agent and then centrifuged.  Spores are very dense compared to the mushroom tissue, bacteria and other impurities, so they separate nicely into a pellet at the bottom of the vial.  Remove the top layer and repeat with a few more washings and centrifuging and you have nearly pure spores when you are done.  This is dried and used as needed.

It is somewhat complicated and not practical for the home experimenter but it does scale up well for the production of commercial quantities of PE spores.  This method can also be used with regular cubensis caps after normal printing.  There are a surprisingly large amount of spores clinging to gills.  Swabbing is easier and works just as well for archiving spore samples.








Has anyone tried this at home? I would love to see this outcome, maybe i see new project in my future.


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OfflineWorkmanV
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Re: How are commercial PE syringes made? [Re: john_conner] * 4
    #25151287 - 04/19/18 05:11 PM (5 years, 9 months ago)

I mentioned this before, but since it isn't in my original post, it often doesn't get added.

You don't put the chunks of mushroom tissue in the centrifuge. You strain out the large pieces with a screen (paint strainer works nicely) adding additional water to wash off any clinging spores. Allow the spore laden wash water to settle for 10 minutes or so (clear glass vessel recommended) and carefully pour off the top water until you see you are starting to lose spores, then add the remaining solution to the tubes for spinning.


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: Workman]
    #25151295 - 04/19/18 05:15 PM (5 years, 9 months ago)

Cool!! when i have some extra time im going to give this a try. I found a few thread on it but nobody has completed it.


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InvisiblemushboyMDiscord
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Re: How are commercial PE syringes made? [Re: john_conner]
    #25151303 - 04/19/18 05:20 PM (5 years, 9 months ago)

:hi:

working man!


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Offlineverum subsequentis
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Re: How are commercial PE syringes made? [Re: mushboy]
    #25151582 - 04/19/18 06:56 PM (5 years, 9 months ago)

i just take a spore laden swab and stir it around in sterile water. then i'll suck and shoot the water passed the swab head a couple of times. works for me:shrug:


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InvisibleShroomyToons
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Re: How are commercial PE syringes made? [Re: verum subsequentis]
    #25151685 - 04/19/18 07:36 PM (5 years, 9 months ago)

Right on, Verum. Why use a machine gun when a rubber band gun will do the trick...:smile:


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OfflineAsuraS
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Re: How are commercial PE syringes made? [Re: Workman]
    #25151729 - 04/19/18 08:00 PM (5 years, 9 months ago)

Quote:

Workman said:
I mentioned this before, but since it isn't in my original post, it often doesn't get added.

You don't put the chunks of mushroom tissue in the centrifuge. You strain out the large pieces with a screen (paint strainer works nicely) adding additional water to wash off any clinging spores. Allow the spore laden wash water to settle for 10 minutes or so (clear glass vessel recommended) and carefully pour off the top water until you see you are starting to lose spores, then add the remaining solution to the tubes for spinning.




If you have some pics or a video of the process, I bet a lot of people would like to see it. This is really fascinating. Just sayin :shrug:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: verum subsequentis] * 1
    #25151738 - 04/19/18 08:08 PM (5 years, 9 months ago)

Quote:

verum subsequentis said:
i just take a spore laden swab and stir it around in sterile water. then i'll suck and shoot the water passed the swab head a couple of times. works for me:shrug:




I could do that oooooor i could over complicate the hell out of it, waste a lot of time and energy to find out that it wont work anyways, yeah i'll do that!:rofl:


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Invisiblesandman420
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Re: How are commercial PE syringes made? [Re: john_conner] * 1
    #25152827 - 04/20/18 08:47 AM (5 years, 9 months ago)

Thanks workman. Can you tell me what speed you use to  spin? 4000 rpm in a 50cc tube good?


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OfflineWorkmanV
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Re: How are commercial PE syringes made? [Re: sandman420] * 2
    #25160012 - 04/23/18 01:53 PM (5 years, 9 months ago)

3000 to 4000 should be plenty.


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InvisibleYesum
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Re: How are commercial PE syringes made? [Re: Workman]
    #25164023 - 04/25/18 08:22 AM (5 years, 8 months ago)

Nothing to add. Just wanna say Ty for a super cool var.:mushroom2:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: Yesum] * 1
    #25164184 - 04/25/18 09:50 AM (5 years, 8 months ago)

got my centrifuge tubes in today, gotta start building the actual centrifuge now. Tubes are much lager then i expected but i will make it work.



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OfflineFishLevelMidnight
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Re: How are commercial PE syringes made? [Re: john_conner]
    #25164377 - 04/25/18 11:43 AM (5 years, 8 months ago)

Maybe I should try this with APE. I'd love to get a pure spore sample and see the proportion of albino to wild type spores.

I got a bunch of dried fruits, so I would crush the caps in some water and then strain through a 22 uM filter into a centrifuge tube. Then spin that to pellet the spores? Then wash the pellet a few times.

Not sure if going to 22 microns with all of the cap material would be viable or not.

What is the wetting agent you are adding workman, is it some sort of detergent like Tween-20 or Triton X-100 to help dissociate the spores?


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25164742 - 04/25/18 03:15 PM (5 years, 8 months ago)

In one of RR videos he talks about using sex lube to help keep the spores from sticking together I wonder if that would help seperate the debris?


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OfflineAsuraS
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Re: How are commercial PE syringes made? [Re: john_conner] * 1
    #25164973 - 04/25/18 04:52 PM (5 years, 8 months ago)

Quote:

john_conner said:
got my centrifuge tubes in today, gotta start building the actual centrifuge now. Tubes are much lager then i expected but i will make it work.






Are you going to make that salad spinner centrifuge?


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: Asura]
    #25165355 - 04/25/18 07:18 PM (5 years, 8 months ago)

Asura I'm not sure yet, I found I few 3d printable centrifuges but to be honest i don't know if I trust printed abs plastic whipping around at thousands of RPM's so I might go with solid plastic or aluminium. I'm still doing some research so if anyone comes across anything interesting please post a link.

I found plans for a centrifuge using a rheostat, ESC and a brushless motor and if you wanted to get real fancy theres some coding for a arduino but that has a max rpm of 3-4k and I'm not sure if it could handle the large tubes I have. We will see in the near future


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Re: How are commercial PE syringes made? [Re: Workman]
    #25165612 - 04/25/18 09:05 PM (5 years, 8 months ago)

This is some advanced shit right here!

Cubenis scientists! Well down ya'll.

:justastonishing:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: LogicaL Chaos]
    #25165620 - 04/25/18 09:09 PM (5 years, 8 months ago)

never said i knew what the hell i was doing hahah google is our friend!


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OfflineLogicaL ChaosM
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Re: How are commercial PE syringes made? [Re: john_conner]
    #25165632 - 04/25/18 09:15 PM (5 years, 8 months ago)

Sounds like u got a good handle of the mechanics of it.

Good luck! :popcorn:

I wonder if u could use like a mini vacuum to suck the spores out of the gills without sucking up dirty air :strokebeard:


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight] * 1
    #25165651 - 04/25/18 09:20 PM (5 years, 8 months ago)

:popcorn:


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Re: How are commercial PE syringes made? [Re: LogicaL Chaos] * 1
    #25165669 - 04/25/18 09:25 PM (5 years, 8 months ago)

should be fun! or completely fail and i cry myself to sleep, either or.

Quote:

LogicaL Chaos said:
I wonder if u could use like a mini vacuum to suck the spores out of the gills without sucking up dirty air :strokebeard:



now that would take some brain storming!


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25165670 - 04/25/18 09:26 PM (5 years, 8 months ago)

Can we get an engineer in this bitch?!


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25166074 - 04/26/18 04:15 AM (5 years, 8 months ago)

Quote:

john_conner said:
never said i knew what the hell i was doing hahah google is our friend!




Not so sure about that. But ok.


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Re: How are commercial PE syringes made? [Re: Yesum]
    #25166082 - 04/26/18 04:26 AM (5 years, 8 months ago)

:takingnotes::popcorn:


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Re: How are commercial PE syringes made? [Re: catnip40]
    #25176024 - 04/30/18 05:18 PM (5 years, 8 months ago)

Okay i decided to go with the smaller centrifuge tubes, the larger ones are just too large, i think the smaller ones will be easier to manage. I have a few things to tighten up in the garage before i can break out my 3D printer. I opt'ed to go with a 3D printed centrifuge built with a old hard drive.

1.5ml tubes




   
It's going to look something like this. People are ripping the hard drives all apart and programming with an arduino, I am not going that fancy for my first one. My plan is to make it on or off (7200rpm) with the existing board for now, then maybe down the road make it more fancy. Either way this should be fun.





After i bought the centrifuge tubes I realized that they are not autoclavable and they do not come sterilized, will that be an issue?

Also should there be a new thread for this or is okay to keep going on this one?


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25176161 - 04/30/18 06:20 PM (5 years, 8 months ago)

Id stsrt my own or ask the thread owner


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Re: How are commercial PE syringes made? [Re: Yesum]
    #25176180 - 04/30/18 06:31 PM (5 years, 8 months ago)

If you want to share you trials here I have no problem with that.


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Re: How are commercial PE syringes made? [Re: sandman420]
    #25176219 - 04/30/18 06:53 PM (5 years, 8 months ago)

Guys... look into something called a dremelfuge.
They hold 6 x 3ml tubes, but are easy to print on a 3D printer, and they attach easily and cheaply to a dremel.

https://www.thingiverse.com/thing:1483
https://github.com/cathalgarvey/dremelfuge

Don't waste any time with the one with the chuck for drills. I tried it, it's total garbage. Got to be the one for the dremel.

if anyone prints me a few (they break! kinda dangerous...) I'll send you some centrifuge tubes for free. Best plastic is one that is very hard; softer plastics will stretch and cause wobbling issues which are less then ideal.

I want to play around with protoplast fusion, and a few processes, such as extracting the chitinase enzymes from snail guts, one of these are needed. If you can print one, I'd pay for it if it isn't ridiculously costly (like most 3D items are, not going to pay $50 to get a plastic item when I can get a used centrifuge that is even better for a bit more).


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Re: How are commercial PE syringes made? [Re: Burbles]
    #25176581 - 04/30/18 09:34 PM (5 years, 8 months ago)

I saw the dremelfuge before, definitely a simple approach but i would like something I don't have to hold as its ripping around at 10's of thousands of rpms haha. I would feel more comfortable if it blew up somewhere else other then my hand.:rofl:


Burbles, is this the exact one you want? https://www.thingiverse.com/thing:1483


if you don't mind waiting a couple of weeks i can print it for you. I haven't ran the 3D printing in over a year so it needs a really good tune up and re calibrating, so its going to be some time before its up and running good again, plus all the shit i have to clean up in the garage.


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Re: How are commercial PE syringes made? [Re: sandman420]
    #25176586 - 04/30/18 09:37 PM (5 years, 8 months ago)

Quote:

sandman420 said:
If you want to share you trials here I have no problem with that.



cool, thanks:mushroom2:


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25177940 - 05/01/18 01:49 PM (5 years, 8 months ago)

Yes that is the correct one. I had one before, and its fine: just use a ring stand and a plastic bucket to attach the dremel onto it. Then you can plug it in far away for safety. :smile:

Super ghetto science equiptment, but it works and it is cheap. In fact, it works *really* well - IIRC at the max speed of the dremel, each gram has a weight of 50 lbs. it's like a super centrifuge. At that speed though, the plastic is *very* tempted to fall apart for obvious reasons.


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Re: How are commercial PE syringes made? [Re: Burbles]
    #25189932 - 05/07/18 03:49 PM (5 years, 8 months ago)

Finally got around to breaking out the 3D printer just to find another set back. One of the linear bearing guides has started to break apart, I'm going to epoxy it back together and hopefully it will hold together long enough to print a replacement part. So once this is fixed i can hopefully get back on track and begin printing the centrifuge!




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Re: How are commercial PE syringes made? [Re: john_conner]
    #25189979 - 05/07/18 04:17 PM (5 years, 8 months ago)

Print a replacement part first after the temporary fix


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Re: How are commercial PE syringes made? [Re: bodhisatta]
    #25211259 - 05/17/18 08:09 PM (5 years, 8 months ago)

So I am way be hide on this project because like most of us, this is an extremely busy time. So, whenever I get some free time at work I try to chip away at it. I was able to epoxy the broken piece back together on the 3D printer but haven't been able to print anything yet(hopefully soon).


I started to tear down a hard drive today, but ran into an issue. As I am trying to keep it as simple as possible and avoid any kind of programming. The issue here is, when I add power to the hard drive, about 30 seconds to a minute the motor stops spinning. I'm guessing its programmed to idle when it not reading/writing. Does anyone have any suggestions around this?

Hard drive with power supply





tear down process






The more I think about the hard drive centrifuge the more I think I may have to abandon it and figure out another way. The dremel centrifuge is cool but like I said before, I don't want to be holding this creation as its ripping around at thousands of rpm's. Maybe someone can think of a way to mount the dremel so I don't have to hold it, I'm open to any and all suggestions.




This is defiantly simple, and seems to be effective, but on the other hand could potentially be dangerous




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Re: How are commercial PE syringes made? [Re: john_conner]
    #25211467 - 05/17/18 09:58 PM (5 years, 8 months ago)

Drill a small hole in the lid of a tub, put centrifuge inside the tub? :shrug:


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Re: How are commercial PE syringes made? [Re: Asura] * 1
    #25211888 - 05/18/18 06:14 AM (5 years, 8 months ago)

Why not go with the paper and string centrifuge I posted?

Wayyyyyy easier and safer and I think it would still work for separating spores from the supernatant


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25213314 - 05/18/18 06:59 PM (5 years, 8 months ago)

So i came across a Dremel drill press stand that i will just make out of scrap wood i have laying around. The string centrifuge is cool but i would like to be able to do multiple tubes at a time and be able to let it run with out me having to do any work. Again i hope i can get to my printer for real this time and get some shit done!
Thanks for the input guys



Like i said I'm just going to build a very simplified version of this with some kind of enclosure just in case of failure.



Dremel Drill Press Stand



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Re: How are commercial PE syringes made? [Re: john_conner]
    #25213980 - 05/19/18 05:38 AM (5 years, 8 months ago)

Just found this.



I'm gonna try this when i find a small tube.
I'll super-duper-duper-glue it to a BIG button.
It'll be for 1 tube but realistically how often are you gonna use it?


Edited by CHOOS (05/19/18 05:42 AM)


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Re: How are commercial PE syringes made? [Re: CHOOS]
    #25276384 - 06/18/18 11:17 AM (5 years, 7 months ago)

Hello everybody long time!
I just wanted to do an update, I did not abandon this project but I have ran into alot of personal isses that have kept me from it.

First I ran into a big health issue that held me up for a while than alot of things went wrong around the house. Finally getting back on track and hopefully start back up with tbis project.

I got the printer out just to find out it has alot of issues itself and need repair. I'm putting the printer on the backburner for now. I will continue by making the parts by hand. Yes I realize there are simpler ways to make this  but I want to try it the way I have it envisioned in my head.

I started a new job so how I dont have access to lathes mills and waterjet anymore so the finished project may not be a pretty as I wanted it to be but hey its a proto type so fuck it.

I'm hoping to get a start on it soon and I will keep everyone posted on my new progress.


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25276389 - 06/18/18 11:20 AM (5 years, 7 months ago)

I’m hoping to do a trial run with some normal cube spores, then try and get spores from dried APE caps.
Summer is a little slower in the lab, so one evening I’ll bring my materials in and see what I can do. Hoping in the next month, but I will update with pics and my method if people are interested.

Good luck brother! Glad things are back on track, stay well.


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25276482 - 06/18/18 12:40 PM (5 years, 7 months ago)

Excited to see how this progresses for you guys. Super cool ideas!


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25276559 - 06/18/18 01:20 PM (5 years, 7 months ago)

Quote:

john_conner said:
Hello everybody long time!
I just wanted to do an update, I did not abandon this project but I have ran into alot of personal isses that have kept me from it.

First I ran into a big health issue that held me up for a while than alot of things went wrong around the house. Finally getting back on track and hopefully start back up with tbis project.

I got the printer out just to find out it has alot of issues itself and need repair. I'm putting the printer on the backburner for now. I will continue by making the parts by hand. Yes I realize there are simpler ways to make this  but I want to try it the way I have it envisioned in my head.

I started a new job so how I dont have access to lathes mills and waterjet anymore so the finished project may not be a pretty as I wanted it to be but hey its a proto type so fuck it.

I'm hoping to get a start on it soon and I will keep everyone posted on my new progress.




We're all glad that you recovered from the health issues! life just gets in the way all the time to show us whos boss.

these are cheap and might work with this project.? centrifuge tube adapter


https://www.labsupplyoutlaws.com/fisher-centrific-model-225-228-benchtop-centrifuge--tube-adapter.htm

EDIT - also wanted to add that the WD-HDD should have a few pins exposed for a jumper. that should turn on/off power save features for the disc spinup.


Edited by k5hd2y (06/18/18 01:32 PM)


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: k5hd2y]
    #25277202 - 06/18/18 06:00 PM (5 years, 7 months ago)

k5hd2yS hey thank you for the great tips! never even thought about those jumpers! that adapter is great but im too cheap haha.


Yes its a little retarded....alot of retarded right now, but until i can get my printer fixed or get my hand on real tools this going to have to make do for now. this is all made from crap just laying around.

i just really wanted to test out the electronic and get them to work, and they do! I have video but i don't think i can embed video on here? (if i can please let me know)I dont want to upload it to you tube.

anyways i think it will work just find for now. Please make comments i want to hear how to make it better and what/how to test first!!!


Well here she is the first proto type!

I call her the spore spinner 3000 V1.0 haha



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Re: How are commercial PE syringes made? [Re: john_conner]
    #25277348 - 06/18/18 07:07 PM (5 years, 7 months ago)

A nice test would be some blood. See if you can separate it.

Easy/cheap to get too :P


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25278371 - 06/19/18 08:10 AM (5 years, 7 months ago)

Haha maybe!

So I was thinking about what water I should use to make these spore syringes. I'm not sure what I should go with. I can get de ionzed water from work or will distilled work just as well?

Hoping to do a test run here in a day or two. I do have a concern that the angle is not great enough for the centrifuge tubes, but like I said I'm going to do a test run and see what happens.

Ill keep you all posted and will probably send some syringes out for people to test to see what results they get.


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25278392 - 06/19/18 08:23 AM (5 years, 7 months ago)

I would use distilled or even regular tap that is PCed.

Good luck man!

How are you filtering? I plan to crush up some dried caps in water, then strain through cheesecloth, then strain through 22 uM filter into 1.5 mL tube (or 15 mL/50 mL conical depending on amount I get) for centrifugation. I will use PCed water to wash the spore pellet 2-3 X before putting to a syringe or drying on foil.


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight] * 1
    #25278407 - 06/19/18 08:34 AM (5 years, 7 months ago)

Distilled for syringes. You don't want spores to germinate then mycelium to lose vigor with nothing to eat.


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Re: How are commercial PE syringes made? [Re: bodhisatta]
    #25281806 - 06/20/18 07:34 PM (5 years, 7 months ago)

Okay did a little test run today. Being it was a test i just used filtered water not distilled, and did everything out in the open, again just testing out the "prototype".


I cut chunks off of sad looking GT cap, submerged it in the water, then stirred it around.




Then I took a small piece of a cotton ball and dropped it in the solution to act as a filter. That set up worked damn well, and sucked it up with a syringe.



Cotton ball in the solution at the top about 11 o'clock position



Syringe with filtered spore solution






Solution in vial



For the centrifuge I re cut the holes to be on a steeper angle, if you look at the previous pictures you can see the liquid is more on the side of the vial verses the bottom(the pictures were taken as the centrifuge was spinning).






I did a run at half speed for 5 minutes and this is the results!











    All in all this is really cool, I still have a ways to go and a lot to still research to do it properly. I'm still having issues with my 3D printer which really sucks, but when its up and running again I'm going to print out a nice enclosure to house everything.

    I do have a few questions, when I run a batch for real should I be doing everything in front of a flow hood? does the distilled water need to be sterilized and do I have to make a SHIP for the distilled water? I'm guessing I need to be a sterile as I can but I'll test out a few methods and see my results.

  Once I figure this stuff out I will need some volunteers to test this out for me, I will cover shipping as long as you post results.


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25281890 - 06/20/18 08:11 PM (5 years, 7 months ago)

nice its pelletized - that is diy science!:rockon:


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Re: How are commercial PE syringes made? [Re: k5hd2y] * 1
    #25282623 - 06/21/18 06:54 AM (5 years, 7 months ago)

Heck yea way to go john_conner, that's waht I'm talking about. I think starting with sterile water and doing as much as possible in front of a hood would be a good idea.


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Re: How are commercial PE syringes made? [Re: sandman420]
    #25282846 - 06/21/18 08:55 AM (5 years, 7 months ago)

Was it a fresh cap?

I want to dry with dry, not sure if you can separate from feuitbody matter once it is dry and smashes up?


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight] * 1
    #25282856 - 06/21/18 09:02 AM (5 years, 7 months ago)

Quote:

fishermansjc said:
Was it a fresh cap?

I want to dry with dry, not sure if you can separate from feuitbody matter once it is dry and smashes up?



I believe Workman uses dried caps in his set up


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Re: How are commercial PE syringes made? [Re: Germs]
    #25282918 - 06/21/18 09:43 AM (5 years, 7 months ago)

Ha! that's actually pretty damn smart?! I'm stealing this idea :P


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Re: How are commercial PE syringes made? [Re: Throwawaygrower]
    #25282942 - 06/21/18 09:54 AM (5 years, 7 months ago)

[Quote]
Was it a fresh cap?

I want to dry with dry, not sure if you can separate from feuitbody matter once it is dry and smashes up?





Its was kind of fresh but I will sure try a dried cap as well, though I don't know if the spores are viable once they are dehydrated might get too hot, maybe someone has the answer to that.


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25282968 - 06/21/18 10:03 AM (5 years, 7 months ago)

I doubt it, curious to find out though!


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25284232 - 06/21/18 09:02 PM (5 years, 7 months ago)

That is a good question about the spores remaining viable after a dehydrator run with 160 degree heat.


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Re: How are commercial PE syringes made? [Re: sandman420]
    #25289356 - 06/24/18 02:02 PM (5 years, 7 months ago)

well I have the printer some what running still a few thinks to tweak, having the issue most people do with printing, shit peeling off the table.
The part should still be usable but I'm going to try and fix the issue before printing anymore parts. I'll probably re print this part as well




Does anyone know if shipping the gills contaning spores is a no go? I dont have any PE to test out here.

Also has anyone by any chance found anymore out about what temps spores can handle?


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25289366 - 06/24/18 02:10 PM (5 years, 7 months ago)

Yeah man, don't open yourself to that risk. In the usa only spores! Store print on foil or a syringe etc is safe to do.


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Re: How are commercial PE syringes made? [Re: k5hd2y]
    #25289368 - 06/24/18 02:12 PM (5 years, 7 months ago)

They can handle a lot. I think 160 they should still work.


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Re: How are commercial PE syringes made? [Re: k5hd2y] * 1
    #25291160 - 06/25/18 03:05 PM (5 years, 7 months ago)

I made up my first syringes today.

Took some B+ caps from January and put into a petridish with some autoclave dh2o. Broke up with a scalpel because I forgot to break them up before adding to the water (doh)

Moved things around a bunch. Forgot anything for straining so I drew the liquid right through a 22 uM filter into a syringe, got about 30 ml with 3 normal sized caps in there.

Put the filtered liquid into 50 mL comical and centrifuge at 4200 rpm for 10 min. Remove supernatant add 20 mL water. Vortex and the centrifuge as before. I did 3 of these washes.

Added 25 mL to final pellet. Vortex to resuspend and then aspirate into 2 10 mL syringes and cap. This concentration is not super dense, but enough spores such that they were visible:


I did this all in front of a flowhood or under a flame.

Not expecting it to be clean. Hoping for clean enough and for germination (these spores were dehydrated at 160F ~24 hours before packaging, then another 24 hours after 5 months storage). Putting some spores to agar tonight and later this week if those don’t germinate.

I’ll post results here.

If I works I may make up 4-6, 3 mL syringes of B+ if others want to test it out (especially interested in those going straight to grain or pf Tek as I am testing on agar.


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InvisibleGerms
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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25291323 - 06/25/18 04:55 PM (5 years, 7 months ago)

Quote:

fishermansjc said:
I made up my first syringes today.

Took some B+ caps from January and put into a petridish with some autoclave dh2o. Broke up with a scalpel because I forgot to break them up before adding to the water (doh)

Moved things around a bunch. Forgot anything for straining so I drew the liquid right through a 22 uM filter into a syringe, got about 30 ml with 3 normal sized caps in there.

Put the filtered liquid into 50 mL comical and centrifuge at 4200 rpm for 10 min. Remove supernatant add 20 mL water. Vortex and the centrifuge as before. I did 3 of these washes.

Added 25 mL to final pellet. Vortex to resuspend and then aspirate into 2 10 mL syringes and cap. This concentration is not super dense, but enough spores such that they were visible:


I did this all in front of a flowhood or under a flame.

Not expecting it to be clean. Hoping for clean enough and for germination (these spores were dehydrated at 160F ~24 hours before packaging, then another 24 hours after 5 months storage). Putting some spores to agar tonight and later this week if those don’t germinate.

I’ll post results here.

If I works I may make up 4-6, 3 mL syringes of B+ if others want to test it out (especially interested in those going straight to grain or pf Tek as I am testing on agar.



:camping:
Spores 2 grain :rockon:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25292588 - 06/26/18 08:09 AM (5 years, 6 months ago)

Nice work man! Cant wait to see results


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OfflineFishLevelMidnight
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Re: How are commercial PE syringes made? [Re: john_conner]
    #25292610 - 06/26/18 08:28 AM (5 years, 6 months ago)

How'd the cotton work for you?

Did you just put it in the water, then suck the water into the syringe through the cotton (eliminating some of the larger chunks)?

I might try this next time (I'm going to grind up like 5 g of B+ caps and do this a little more legit, make a bunch of small-medium syringes for giveaway) and sterilize the cotton before hand.

If things work out and I get germ on my plates, I am decapitating all my dried APE and seeing what I can get out of them. I want for crosses and to run generations/trades/giveawats and to see some albino spores, so I'll post a microscope pic later.


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25292667 - 06/26/18 09:03 AM (5 years, 6 months ago)

I'm probably talking out my arse here but could you not just rig up a small battery drill to act as a centrifuge? Most people these days have a drill at home for diy, then all you would need is maybe some threaded steel bar a few nuts and some kind of adapter like john_conners to hold the vials in place while spinning. I know it's not really set and forget but it could work, assuming the rpm is high enough.

Edit- Found this online, this looks a bit fancier than what I was imagining.


Edited by JADO (06/26/18 09:07 AM)


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: JADO]
    #25293798 - 06/26/18 08:50 PM (5 years, 6 months ago)

Fishermansjc-for filtering the slop of spores and mushroom debris it worked great. I haven't ran a test all the way through yet, I finally have my printer up and running properly so I'm focused on printng out all the centrifuge parts now. Once its done I will run a test all the way through.

Jado- I owe burbles a dremel fuge once I'm done printing my contraption I will print a few of those out, let me know if you want one.

Burbles- didn't forget about you, like I said once I'm done mine I will print you out that dremel fuge


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25294257 - 06/27/18 05:29 AM (5 years, 6 months ago)

Quote:

john_conner said:
Fishermansjc-for filtering the slop of spores and mushroom debris it worked great. I haven't ran a test all the way through yet, I finally have my printer up and running properly so I'm focused on printng out all the centrifuge parts now. Once its done I will run a test all the way through.

Jado- I owe burbles a dremel fuge once I'm done printing my contraption I will print a few of those out, let me know if you want one.

Burbles- didn't forget about you, like I said once I'm done mine I will print you out that dremel fuge




John, I'm totally interested too man! when/if you get the time. :syringe::rocket:


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25295598 - 06/27/18 06:10 PM (5 years, 6 months ago)

Quote:

john_conner said:
Fishermansjc-for filtering the slop of spores and mushroom debris it worked great. I haven't ran a test all the way through yet, I finally have my printer up and running properly so I'm focused on printng out all the centrifuge parts now. Once its done I will run a test all the way through.

Jado- I owe burbles a dremel fuge once I'm done printing my contraption I will print a few of those out, let me know if you want one.

Burbles- didn't forget about you, like I said once I'm done mine I will print you out that dremel fuge




It'd be wasted on me, I've no plans to grow PE just yet but thanks for the offer :thumbup:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: JADO]
    #25303924 - 07/02/18 06:59 AM (5 years, 6 months ago)

Just an update:



Almost finished here and I can finally do a test run all the way through






After I'm done this I will start on the dremel fudge for you guys

:cheers:


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25304090 - 07/02/18 08:52 AM (5 years, 6 months ago)

Quote:

john_conner said:
Just an update:



Almost finished here and I can finally do a test run all the way through






After I'm done this I will start on the dremel fudge for you guys

:cheers:




Jesus.  :thedude:


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Re: How are commercial PE syringes made? [Re: elasticaltiger]
    #25304112 - 07/02/18 09:00 AM (5 years, 6 months ago)

An update on my end.

Of the 2 plates I tried with the spores taken from dried caps (2X at 160F actually) one got super bacterial and the other is showing no growth at all. I think the bacteria came in from shaking the plate later on and not from the syringe, as the other plate shows nothing at all.

I'll follow it a bit further; however, it seems that spores dried at 160F are not viable. I plan to dry some batches of fruit at a lower temp to see if I can keep the integrity of the spores but also dry the fruits properly. I was thinking 100F to start, any suggestions?


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25305010 - 07/02/18 05:35 PM (5 years, 6 months ago)

fishermansjc great work and thanks for the info! how did you wash the spore between each cycle? i was trying to think of the best way to do this.

I finally fished my centrifuge today hope to get a test run in asap.






Still some polishing up to do but you get the point.

Now i will start on the dremel fuges i promised and may go again at the hard drive centrifuge


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25305273 - 07/02/18 08:06 PM (5 years, 6 months ago)

After collecting them the first time through the 22 uM filter I spun them down. Then I used PCed water to wash them (I dumped the supernatant, then added 20 mL, I was doing this in a 50 mL centrifuge tube) by vortexing to break up the pellet. Then I spun down and removed super and repeated 3X. Then added 25 mL and drew up into the two syringes I posted.

Anyone know the thermal tolerance of cubensis spores?


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25307867 - 07/04/18 09:07 AM (5 years, 6 months ago)

okay people I finished the dremel fuges there are two different kinds here. There is one that will go into a drill chuck and one that goes to a dremel.

They are up for grabs, of course k5hd2yS and BurblesV get first dibs, but after that let me know who wants one. I printed four and two are spoken for, so simple math two are left.

I will cover shipping but all i ask for in return is that, if someone can find me some PE spores i would really appreciate it, but if not then no biggie.




Disclaimer:
I have not tested these items out, proceeded at your own risk. I am not responsible for any injury that may happen. Use caution, safety glasses and common sense please.


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Re: How are commercial PE syringes made? [Re: john_conner] * 1
    #25308470 - 07/04/18 03:05 PM (5 years, 6 months ago)

I bet if you keep it under 110 on the dehydrator you will be good.


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Re: How are commercial PE syringes made? [Re: sandman420]
    #25310361 - 07/05/18 05:33 PM (5 years, 6 months ago)

So i lost all of my emails so who ever wanted a dremel fuge will have to re email me because i dont remember who asked for them.
Sorry people


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25310368 - 07/05/18 05:37 PM (5 years, 6 months ago)

They look good brother!


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: Bry Bry]
    #25317377 - 07/09/18 07:20 PM (5 years, 6 months ago)

bry bry - thanks!

Did i quick test run today i wanted to see how lazy you can be with these things. All work was done in the open no flow hood, SAB or gloves, cleaned twice and ran for 3 min per cycle hopefully get them on agar tomorrow to see results!

had a handful of Martinique that went for too long so i just put the caps in distilled water and coarsely filtered through cheese cloth. I let it sit for an hour or so to let the spores settle to the bottom so i could get denser amount in each tube.



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Re: How are commercial PE syringes made? [Re: john_conner]
    #25317648 - 07/09/18 10:28 PM (5 years, 6 months ago)

Very cool john.


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Re: How are commercial PE syringes made? [Re: filthyknees]
    #25317677 - 07/09/18 10:52 PM (5 years, 6 months ago)

Good luck john. Lets see how clean they come out.


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Re: How are commercial PE syringes made? [Re: john_conner]
    #25320373 - 07/11/18 10:25 AM (5 years, 6 months ago)

keep the updates coming:thumbup:


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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: k5hd2y]
    #25327468 - 07/15/18 11:02 AM (5 years, 6 months ago)

Well the first test run went pretty much as expected, all contaimed out, kind of knew that would happen but hey sometimes you get lucky.
:mushroom:

I ran into another issue, i believe my hepa module (fake flowhood) is a cause of alot of contaims, Ive been having massive issues latley, lost 5 out of 6 shoe boxes. the odd thing is that when i do agar dishes with the hepa module they come out fine, so i have to fine tune some things to figure out whats going on. Unless i can figure out how to do it in a SAB, which the centrifuge defeats the purpose of a SAB, Im kind of SOL, Im still going to try it though.:shrug:

:mushroom2:

There are still three dremel fuges left up for grabs, Two drill chuck versions and one dremel version. :pm: if you would like one.



Heres the agar dishes with the first five spores syrgines that i ran




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Invisiblejohn_conner

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Re: How are commercial PE syringes made? [Re: john_conner]
    #25364899 - 08/03/18 05:22 PM (5 years, 5 months ago)

okay people im checking out for awhile good luck to everyone

cya


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OfflineFishLevelMidnight
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Re: How are commercial PE syringes made? [Re: john_conner]
    #25365031 - 08/03/18 06:44 PM (5 years, 5 months ago)

Hope all is well brother. Good luck to you


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Re: How are commercial PE syringes made? [Re: FishLevelMidnight]
    #25365228 - 08/03/18 08:18 PM (5 years, 5 months ago)

Yea brother thanks for all the efforts. Good stuff. Be well.


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