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GalliumPE
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Registered: 10/11/17
Posts: 330
Loc: The Virgo Supercluster
Last seen: 2 years, 17 days
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How to germinate dirty spore swab.
#25140006 - 04/15/18 12:53 AM (6 years, 8 months ago) |
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I have 2 ape swabs that I have been trying to germinate and each one grows bacteria before the spores have a chance to germinate. I tried the swipping method and I even tried to use a sterile scalple to pull off sections of the swab to put on agar. All 4 petri dishes are growing the same bacteria leaving the spores no fighting chance. Is there any way to force them to germinate or am I just sol and the swabs are too dirty to work with? I know its not the petri dishes because all my other spores germinate just fine with no contaminates from the same batch of agar.
Oh I should mention that 2 APE petris are %4 MEA and the other 2 APE petris are %2 MEA.




EDIT 4/19/18: So I'm glad i kept the plates. One of them germinated and im going to do a transfer now. Thanks for the help and suggestions everyone.

The APE Gods have come to my aid!
Edited by GalliumPE (04/19/18 09:09 PM)
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elasticaltiger
Like Tigers in Coitus




Registered: 06/24/13
Posts: 8,438
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Re: How to germinate dirty spore swab. [Re: GalliumPE]
#25140015 - 04/15/18 01:02 AM (6 years, 8 months ago) |
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Nah forget it. You've already tried everything and it sounds like the bacteria highly outnumbers any spores.
If you had said there was definitely mycelium growing along with it then it might be worth salvaging but it sounds like you're getting nothing like that so it's time to trash em.
-------------------- First time growing cakes? DON'T make a Shotgun Fruiting Chamber
The Shmuvbox. - The Old TC's Like it
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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Pony
Part-Time Bra



Registered: 11/01/09
Posts: 287
Loc: Oklahoma
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Is that mycelia in the 4th picture? I agree with elastical, but you might be able to transfer from that one to some more agar, until you get a clean sample.
Or this is a real long shot: get you a horse apple, pasteurize it, and shove the swab in it, bury it in your backyard about 1/4 an inch over the top and hope for a miracle this fall. Don’t really have anything to lose.
-------------------- Sacred cows make the best hamburgers.
Edited by Pony (04/15/18 01:15 AM)
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GalliumPE
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Registered: 10/11/17
Posts: 330
Loc: The Virgo Supercluster
Last seen: 2 years, 17 days
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Re: How to germinate dirty spore swab. [Re: Pony]
#25140059 - 04/15/18 01:34 AM (6 years, 8 months ago) |
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No it's just bacteria but looks like mycelium from the picture a little. Guess I should just cough up the money and buy an APE syringe as swabs are hit or miss.
-------------------- I have no special talents. I am only passionately curious - Albert Einstein
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NarkedAt90ft
Deep in the Blue Abyss


Registered: 12/04/13
Posts: 149
Loc: Marianas Trench
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Re: How to germinate dirty spore swab. [Re: GalliumPE] 1
#25140877 - 04/15/18 11:53 AM (6 years, 8 months ago) |
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Would a firmer agar help swing things in favor of the spores?
Or if you don't have access horse apples maybe try direct to grains? My first syringe was horribly bacterial - had the same results as above, I was attempting agar first before grains. I then hit up some grains, jar turned out awful but I opened it up and slowly dissected it to find a little colony of mycelium... it went back to agar and I managed to save it.
Were it me, I'd tweeze off some of those cotton strands and knock up a couple small jam jars of dryer than usual grains and hope for the best. I hate giving up!
Good luck
-------------------- ni·tro·gen nar·co·sis noun - a drowsy state induced by breathing air under higher than atmospheric pressure, for example, in deep-sea diving. Also known as Rapture of The Deep.
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GalliumPE
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Registered: 10/11/17
Posts: 330
Loc: The Virgo Supercluster
Last seen: 2 years, 17 days
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Re: How to germinate dirty spore swab. [Re: NarkedAt90ft]
#25140910 - 04/15/18 12:05 PM (6 years, 8 months ago) |
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I might try that, I got plenty of WBS and lots of half pints from my old cake grows I used to do. I don't want to give up yet and I still see spores on sections of the swap that haven't touched the agar yet.
-------------------- I have no special talents. I am only passionately curious - Albert Einstein
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dextr0
Stranger


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Re: How to germinate dirty spore swab. [Re: NarkedAt90ft]
#25140912 - 04/15/18 12:06 PM (6 years, 8 months ago) |
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Try putting pieces to brf puck. Josex had a little snippet of how he does it but can't find atm
Edit bottom of first post: https://www.shroomery.org/forums/showflat.php/Number/24806569
Edited by dextr0 (04/15/18 12:12 PM)
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LtLurker
Lost Sailor



Registered: 01/03/18
Posts: 7,537
Loc: Borderlands
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Re: How to germinate dirty spore swab. [Re: dextr0]
#25140936 - 04/15/18 12:13 PM (6 years, 8 months ago) |
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https://www.shroomery.org/forums/showflat.php/Number/24740168
Josex's Poke method from the agar into a brf puck.
*Edit, incorrect Josex Link. Dextr0 got it.
Edited by LtLurker (04/15/18 12:17 PM)
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dextr0
Stranger


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Re: How to germinate dirty spore swab. [Re: LtLurker]
#25140945 - 04/15/18 12:14 PM (6 years, 8 months ago) |
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LT...right I looked there too he moved it to the link I posted though...either way good links.
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LtLurker
Lost Sailor



Registered: 01/03/18
Posts: 7,537
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Re: How to germinate dirty spore swab. [Re: dextr0]
#25140948 - 04/15/18 12:16 PM (6 years, 8 months ago) |
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Quote:
dextr0 said: LT...right I looked there too he moved it to the link I posted though...either way good links.
Oh you're right. I didn't look close enough. Good catch.
Late thought. What about letting the plate pin and xfer the pin for cleaner start?
Edited by LtLurker (04/15/18 12:19 PM)
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Pony
Part-Time Bra



Registered: 11/01/09
Posts: 287
Loc: Oklahoma
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Re: How to germinate dirty spore swab. [Re: LtLurker]
#25141326 - 04/15/18 02:52 PM (6 years, 8 months ago) |
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It looks like there’s way too much bacteria, the only part that looks like mycelium is the cotton from the q-top i think. It’s basically worthless, if you don’t have a horse apple, try cardboard l. Probably still won’t work though.
-------------------- Sacred cows make the best hamburgers.
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dextr0
Stranger


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Re: How to germinate dirty spore swab. [Re: Pony]
#25141705 - 04/15/18 06:11 PM (6 years, 8 months ago) |
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What is a horse Apple? And how does that work?
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PsilocyBen17
Pin Pornographer


Registered: 10/20/13
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Re: How to germinate dirty spore swab. [Re: dextr0]
#25141820 - 04/15/18 06:59 PM (6 years, 8 months ago) |
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what is your technique? Tell us what you are doing to knock up the plates. Though it is possible you could have an exceptionally dirty print, most contams come from the cultivator.
Another thing you could try is a hot pour or agar sandwhich method. Pour agar into another dish n let it cool. Once cooled, place the puck of agar ontop of the agar in the other dish, sandwhiching the bacteria and spores between two slabs of agar. This might gives myc a chance to push through.
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GalliumPE
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Registered: 10/11/17
Posts: 330
Loc: The Virgo Supercluster
Last seen: 2 years, 17 days
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Re: How to germinate dirty spore swab. [Re: PsilocyBen17]
#25142062 - 04/15/18 08:45 PM (6 years, 8 months ago) |
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Trust me, its not the cultivator. Have you ever used a swab before? They are notourious for being dirty compared to prints and syringes. Out of 24 poured dishes, 8 reserved to cubes, 2 to pans, 4 to ape, and the rest in storage until further notice. Only the 4 ape petris contaminated. I think that gives you your answer. Thank you for the sandwhich teqnique but imo I don't think it would work for something this contaminated. Sandwhich teqnique is more often used for spores that won't germinate due to older age which isn't the problem I am facing.
I have decided to just let the newer dishes sit a little longer wrapped up and hope for the best. If no miracle happens by the end of the week then I will just trash them and get a syringe instead of swabs.
-------------------- I have no special talents. I am only passionately curious - Albert Einstein
Edited by GalliumPE (04/15/18 08:49 PM)
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Josex
#cheat_code


Registered: 11/13/15
Posts: 8,999
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Re: How to germinate dirty spore swab. [Re: GalliumPE]
#25142112 - 04/15/18 09:10 PM (6 years, 8 months ago) |
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No myc, only bacteria, that's the problem. Don't toss them yet though, you've nothing to lose at this point and maybe some miracle happens and spores manage to germinate. I remember I had a plate like that from a swab and spores managed to grow amidst all the gunk and could salvage it.
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: How to germinate dirty spore swab. [Re: Josex]
#25142270 - 04/15/18 10:48 PM (6 years, 8 months ago) |
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Haha it happens right? I've seen it before too. I've also seen shit literally get covered in bacteria and that's it. Either way I agree it doesn't hurt to keep em around if you are interested in testing your skills at removing mycelium from bacteria
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GalliumPE
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Registered: 10/11/17
Posts: 330
Loc: The Virgo Supercluster
Last seen: 2 years, 17 days
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Re: How to germinate dirty spore swab. [Re: Mad Season]
#25142324 - 04/15/18 11:47 PM (6 years, 8 months ago) |
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Oh yeah if i do get lucky then its going to be a challenge. Thankfully I have plenty of plates and lots of time on my hands.
-------------------- I have no special talents. I am only passionately curious - Albert Einstein
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Josex
#cheat_code


Registered: 11/13/15
Posts: 8,999
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Re: How to germinate dirty spore swab. [Re: GalliumPE]
#25142340 - 04/15/18 11:57 PM (6 years, 8 months ago) |
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No need for a challenge if you eventually get germination, though. Listen carefully, there might be something worth listening to. You got your answer in this thread already.
Edited by Josex (04/16/18 12:40 AM)
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GalliumPE
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Registered: 10/11/17
Posts: 330
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Last seen: 2 years, 17 days
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Re: How to germinate dirty spore swab. [Re: GalliumPE]
#25151902 - 04/19/18 09:10 PM (6 years, 8 months ago) |
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I got lucky and a miracle happened despite all the contamination!
-------------------- I have no special talents. I am only passionately curious - Albert Einstein
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elasticaltiger
Like Tigers in Coitus




Registered: 06/24/13
Posts: 8,438
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Re: How to germinate dirty spore swab. [Re: GalliumPE]
#25152260 - 04/20/18 12:48 AM (6 years, 8 months ago) |
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Was that just a rub on the agar or was that a piece of the cotton?
-------------------- First time growing cakes? DON'T make a Shotgun Fruiting Chamber
The Shmuvbox. - The Old TC's Like it
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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GalliumPE
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Registered: 10/11/17
Posts: 330
Loc: The Virgo Supercluster
Last seen: 2 years, 17 days
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It was a piece of cotton, I carefully did 3 transfers from that spot. It is %100 indeed mycelium after taking a closer look. After letting it grow out and making a few more transfers I should have a viable culture worth saving. From my guess I'd say when I swipped the agar the first time on the other plates some of the agar stuck to the cotton. I believe that contributed to the germination of the spores on the cotton itself which then started taking over the bacteria around the rest of the agar. It is currently winning it looks like against all the bacteria which is pretty amazing imo as I was beginning to think about throwing the plates out.
Edited by GalliumPE (04/20/18 01:39 AM)
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