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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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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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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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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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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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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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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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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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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

EZEKIEL 23:20


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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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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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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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Offlinefunga
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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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Invisiblejohn_conner

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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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OfflineFishLevelMidnight
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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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InvisiblebodhisattaMDiscordReddit
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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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Invisiblesandman420
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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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InvisibleGerms
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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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OfflineFishLevelMidnight
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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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