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 FishLevelMidnight
Aquaman
 
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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 Fish's successful germination from stubborn spores; swab to agar TEK  9
#25162436 - 04/24/18 01:45 PM (5 years, 11 months ago) |
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Hey all!
I have been lucky enough to collect a wide variety of spores during my time here on the shroomery; the good, the bad, and the ugly.
I have had trouble with a few spore swabs from PE getting any germination after swabbing plates, even plates with a low agar content, and came up with this method to help combat it. It is probably too early to call this a TEK just yet, but I am replicating this TEK with old spores and some other varieties that didn't germinate for me and I recommend others to try it as well.
What you will need:
Sterile scissors
Sterile agar plate
Sterile water
Sterile scalpel
Spore swab or sterile swab to make spore swab
The TEK:
1. Position all your items in front of the flowhood or in an SAB.
2. If you are working with a stubborn print, first make a swab from the print. Try to collect the spores to one side.
3. Cut a small square or circle from the center of the agar. It is best to be just slightly larger than the tip of the swab and to not cut all the way through the agar (make a well in the center of the dish).
4. Carefully position the swab tip in the center of the well you just made, then cut it with the scissors such that it falls into the center of the well (preferably spores downward- you can use a sterile scalpel or sterile forceps to position the swab tip as you'd like.
5. Carefully soak the swab tip with sterile water. Having some excess water in the bottom of the well is fine, but you want to eliminate the ability for water to slosh all around the plate.
6. Wrap and let sit agar side down until germination.
7. Proceed with transfers 
Results:
Here is the only photo I've taken thus far from my TEK.
Here is that plate after quite some time (I should have probably cloned that PE pin...)
Pins on plate
I used a PE MS swab that was taken from fruit and wouldn't germination after 3 separate attempts (1 in soft agar). I did the above tek and the entire spore swab is now a puffball of myc, leaving myc in the well for me to transfer from. My swab was shaken loose while traveling with the plate and caused germination in another location of my plate- this could be used to our advantage as cultivators as a colonized swab tip can then be transferred to subsequent dishes to expand upon the germinated myc.
Any tips or questions are appreciated! Anyone who wants to try this out should post results too
Edit-
I have been working a little with CBKs albino and the spores were hard to germinate.
I got a culture, for a nice grow and swabbed.
My first swabbed failed on the plate.
I used another that I swabbed and then left with this Tek and I got germination now
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Edited by FishLevelMidnight (09/02/19 06:49 PM)
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P.Avgvst
Registered: 03/13/16
Posts: 56
Last seen: 4 months, 24 days
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 Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25163176 - 04/24/18 07:37 PM (5 years, 11 months ago) |
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Woah, thats a really good procedure! I’m going to try with some PE spores I have. Thank you for sharing, man
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 Munchauzen
Registered: 06/22/11
Posts: 14,344
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight] 1
#25163265 - 04/24/18 08:26 PM (5 years, 11 months ago) |
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MrRipley
Hola!
Registered: 04/05/14
Posts: 24
Last seen: 5 years, 6 months
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Munchauzen]
#25163404 - 04/24/18 09:56 PM (5 years, 11 months ago) |
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Just what the doctor ordered 
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Haywire
Wetspot Wizard
Registered: 12/29/13
Posts: 1,619
Last seen: 2 hours, 8 minutes
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25163592 - 04/25/18 12:39 AM (5 years, 11 months ago) |
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Quote:
fishermansjc said:
this could be used to our advantage as cultivators as a colonized swab tip can then be transferred to subsequent dishes to expand upon the germinated myc.
Good idea!
would you use the myceliated qtip as a swab or would you just put it on the agar, let the agar colonize and then transfer to a new dish?
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Ciao mamma, guarda come mi diverto
My grows  Outdoor patches
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Haywire]
#25163754 - 04/25/18 04:48 AM (5 years, 11 months ago) |
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Quote:
Haywire said:
Quote:
fishermansjc said:
this could be used to our advantage as cultivators as a colonized swab tip can then be transferred to subsequent dishes to expand upon the germinated myc.
Good idea!
would you use the myceliated qtip as a swab or would you just put it on the agar, let the agar colonize and then transfer to a new dish?
I would do the latter. Use the colonized qtip as if it was a colonized agar wedge. Once you have germ and it spreads to the plate, the qtip can be transferred again. I would imagine that this wouldn’t last too many iterations to protect from contams
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Haywire
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25163796 - 04/25/18 05:31 AM (5 years, 11 months ago) |
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Quote:
fishermansjc said:
I would do the latter. Use the colonized qtip as if it was a colonized agar wedge. Once you have germ and it spreads to the plate, the qtip can be transferred again. I would imagine that this wouldn’t last too many iterations to protect from contams
wouldn't it make more sense then to swipe? swipe 5-10 new plates and be done with it.
then you only handle it once. I'm sure the fragmented mycelium would recover although maybe a bit slower.
I'll have to try it out. I'll post my results here if that's ok?
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Ciao mamma, guarda come mi diverto
My grows Outdoor patches
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Haywire]
#25163808 - 04/25/18 05:45 AM (5 years, 11 months ago) |
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Yeah that sounds great!
I supposed you could grab the colonized swab with some forceps and just streak with it...
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Haywire
Wetspot Wizard
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight] 1
#25163847 - 04/25/18 06:33 AM (5 years, 11 months ago) |
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I'll try it with that pink buffalo print you've sent me.
I'm sure it'll have no problem germinating but I'm more interested in swabbing the colonized qtip anyway.
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Ciao mamma, guarda come mi diverto
My grows Outdoor patches
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CHOOS
Stranger
Registered: 08/05/15
Posts: 105
Last seen: 3 years, 10 months
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Haywire] 1
#25164288 - 04/25/18 10:47 AM (5 years, 11 months ago) |
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Small improvement = fucking genius!
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Haywire
Wetspot Wizard
Registered: 12/29/13
Posts: 1,619
Last seen: 2 hours, 8 minutes
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: CHOOS]
#25168392 - 04/27/18 05:09 AM (5 years, 11 months ago) |
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it's on. I'm using your print (thai pink buffalo and a golden teacher print from funga)
However, I didn't cut out the agar in the middle, didn't really think it was necessary.
I also put some spores straigt on the agar, just to see if there's a difference in germination time.
 
UPDATE:
I got clean germination from both prints (spores only) which means the print are as clean as they can get.
however I have bacterial contamination from the spore swabs. this could mean that either the swabs weren't entirely clean or the sterile water. I'm guessing the water, since I've had it for a while in the lab. I'll repeat the experiment.
from the looks of it, the TPB has less bacteria than the GT.
 
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Ciao mamma, guarda come mi diverto
My grows Outdoor patches
Edited by Haywire (05/03/18 12:57 AM)
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Haywire]
#25168404 - 04/27/18 05:21 AM (5 years, 11 months ago) |
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Fucking love it! Very excited to see results
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AndyHinton
Registered: 12/05/16
Posts: 434
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25168434 - 04/27/18 05:55 AM (5 years, 11 months ago) |
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Hey fisherman, this is a great idea. I may want to try something like it for reviving saline. It gets me thinking about how to adapt gel electrophoresis combs, i.e., pouring plates around an obstacle to make wells in the agar.
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Edited by AndyHinton (04/27/18 06:01 AM)
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: AndyHinton]
#25168483 - 04/27/18 06:31 AM (5 years, 11 months ago) |
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Dang that is a lovely idea!
The only thing that seems difficult is doing so and keeping everything sterile.
I suppose if you had a pp5 piece of something you could insert it in the agar as it was cooling, then remove it under a flowhood and replace with the swab...
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25268864 - 06/14/18 09:30 PM (5 years, 9 months ago) |
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Pins on plate
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Bry Bry
Mental
Registered: 01/18/17
Posts: 575
Loc: Somewhere in time 
Last seen: 3 years, 5 months
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25326600 - 07/14/18 08:01 PM (5 years, 8 months ago) |
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Haven't gotten anything from the swabbing but have 11 transfers from the swab head. Had to do the same with an APE swab. Pretty sure this thread is the reason I remembered to do it.
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Bry Bry]
#25326615 - 07/14/18 08:10 PM (5 years, 8 months ago) |
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That’s a great example of the TEK in action! Check out the swab, all covered in myc, even when a plate swab did nothing.
I hope the PE gets going for you, about to put my last swab to agar myself.
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FishLevelMidnight
Aquaman
Registered: 09/01/17
Posts: 2,328
Last seen: 8 months, 9 days
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25342846 - 07/23/18 06:04 PM (5 years, 8 months ago) |
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I’ll take some more pictures tonight but this has become my go too for stubborn spores.
Really great for PE, PEU and APE; I have a B+ PE cross that I was able to revive from this method
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Bry Bry
Mental
Registered: 01/18/17
Posts: 575
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Last seen: 3 years, 5 months
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: FishLevelMidnight]
#25342870 - 07/23/18 06:14 PM (5 years, 8 months ago) |
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Had to finally toss the swab after getting a few more transfers. Was going bacterial to the point of no return. Worked like a charm tho.
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AlexP
Registered: 06/04/09
Posts: 2,270
Loc: Europe
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Re: Fisherman's successful germination from stubborn spores; swab to agar TEK [Re: Bry Bry]
#25380879 - 08/11/18 03:40 AM (5 years, 8 months ago) |
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"To feel today what one felt yesterday isn't to feel - it's to remember today what was felt yesterday, to be today's living corpse of what yesterday was lived and lost." Fernando Pessoa
"When one creates phantoms for oneself, one puts vampires into the world, and one must nourish these children of a voluntary nightmare with one's blood, one's life, one's intelligence, and one's reason, without ever satisfying them." Eliphas Levi
"The joy of life consists in the exercise of one's energies, continual growth, constant change, the enjoyment of every new experience. To stop means simply to die. The eternal mistake of mankind is to set up an attainable ideal." Aleister Crowley
"The Universe is an artistic catharsis." Artifex Infinitum
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