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OfflineTheApprentice
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Q tips to transfer spores to agar. No germination
    #26388453 - 12/18/19 10:15 AM (4 years, 1 month ago)

It’s been years since my last grow.  Returned to visit my parents recently and found all my old tools, including my print collection.  So I quickly made up some Pasty Plates and used three different cube varieties on several different plates.  Worked in SAB.  It’s been about 13 days now with no visible signs of growth on any of them.  I can’t figure out why none of my plates will germinate.

Any thoughts?  Is it possible the damn spores didn’t transfer at all from the cotton swab?


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OfflineCrackatoa
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Re: Q tips to transfer spores to agar. No germination [Re: TheApprentice]
    #26388460 - 12/18/19 10:19 AM (4 years, 1 month ago)

I've had it happen before. Give a little while longer.


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OfflineeLeSDenes
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Re: Q tips to transfer spores to agar. No germination [Re: Crackatoa]
    #26388517 - 12/18/19 10:51 AM (4 years, 1 month ago)

After two years the germination success rate decreases very rapidly. There are examples that people managed to germinate after 5-10 years but it is very rare and the method of storage is very important too. If you have your spores on foil or glass and stored in the fridge, you have the best chance, paper and swabs and no fridge... astronomically small. 

There are various teks here for old spore germination. The main problem usually is that spores dehydrate and you have to re-hydrate them. You can make your agar a bit more soupy or re-hydrate in sterile distilled water for a bit of time before putting them on agar.


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OfflinecronicrFacebook
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Re: Q tips to transfer spores to agar. No germination [Re: eLeSDenes]
    #26388530 - 12/18/19 10:59 AM (4 years, 1 month ago)

Get new spores noob


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Invisiblegizmo1
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Re: Q tips to transfer spores to agar. No germination [Re: eLeSDenes] * 1
    #26388536 - 12/18/19 11:05 AM (4 years, 1 month ago)

I'd say try scraping a bunch of spores onto the plate instead of using a cotton swab. More spores will give you a better chance of some germination.
Supposedly brf pucks work good for germinating spores. Never tried it myself though.


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OfflineBLINKfan420
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Re: Q tips to transfer spores to agar. No germination [Re: gizmo1]
    #26388644 - 12/18/19 12:19 PM (4 years, 1 month ago)

I make my agar recipe with a little extra water so the plates come out really moist and jelly like.  It seem like my older spores have had a better chance at germinating on the wetter plates than the drier ones.  That's not fact but just my experience. :shrug:

BLiNK


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OfflineCrackatoa
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Re: Q tips to transfer spores to agar. No germination [Re: BLINKfan420]
    #26388649 - 12/18/19 12:22 PM (4 years, 1 month ago)

I've got some LPEU prints if you want one, hit me up.


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Offlinedaysbetween
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Re: Q tips to transfer spores to agar. No germination [Re: Crackatoa]
    #26389045 - 12/18/19 04:02 PM (4 years, 1 month ago)

I'd give 'em at least a month.  Old spores can take a reallly long time if they are still viable.  Only time will tell.

Did you use q-tips to transfer? or am I reading that wrong.

I hope they were sterile swabs and not just qtips.


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OfflineAsuraS
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Re: Q tips to transfer spores to agar. No germination [Re: daysbetween]
    #26389073 - 12/18/19 04:20 PM (4 years, 1 month ago)

I remember you, dude. Welcome back.


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InvisibleDigit
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Re: Q tips to transfer spores to agar. No germination [Re: Asura]
    #26389183 - 12/18/19 05:13 PM (4 years, 1 month ago)

Im in a very similar position right now.
I recently put a bunch of 3 year old spores and cultures onto agar with very limited success so far.
I had a handful of spore syringes that had been in the fridge for about 3 years, a few petris with isolates I made 3 years ago, and a collection of azurescens prints from a friend in Oregon, also about 3 years old.
So far none of the spores have germinated at all. Nothing. It's been over 2 weeks and not even contams are growing on the dishes.
A couple days ago I kinda went nuts and just scraped a massive pile of spores from one of the Azzie prints onto a few dishes. Im hoping that within a few million spores there will be enough viable ones to get a culture going. If that doesn't work I'll try rehydrating them first.
The wedges I transferred from my old petris are finally starting to take off, but it took a long time and I'm hesitant to put all my bets on them at this point.
I always thought that spores stayed viable for many years if refrigerated.
Apparently I was wrong about that.
I finally just ordered a 4 pack of different cube varieties and I'll be comparing them with each other and also with my surviving isolates.
I'm thinking maybe shoe boxes will be a good way to compare several different cultures with each other so I can pick the best ones for further research with full size monos.
Best wishes


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Believe nothing.
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Offlinespiritlands
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Re: Q tips to transfer spores to agar. No germination [Re: gizmo1]
    #26389401 - 12/18/19 07:13 PM (4 years, 1 month ago)

Quote:

gizmo1 said:
I'd say try scraping a bunch of spores onto the plate instead of using a cotton swab. More spores will give you a better chance of some germination.
Supposedly brf pucks work good for germinating spores. Never tried it myself though.



This is what I've been doing with all my old spores as well. I've had some prints germinate no prob but others not at all. The ones that don't germinate in 3 weeks I've been tossing and redoing being generous with the spores. I haven't been doing anything special with the agar either. Most of what I'm working with are 8 years old or more.


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InvisibleBuddaking
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Re: Q tips to transfer spores to agar. No germination [Re: spiritlands]
    #26389510 - 12/18/19 08:20 PM (4 years, 1 month ago)

My old print collection came back to me out of nowhere. Most are 10 yr oldish. Storage last 4 years is my enemy. In a garage exposed to extreme temperature fluctuations.
I took 10 strait to PDA & a dozen others i made syringes with activated charcoal, capped them and am applying pressure with rubber bands in an attempt to force hydrate. Prob a futile attempt but time will tell.


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OfflineCocaineBuffet
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Re: Q tips to transfer spores to agar. No germination [Re: Buddaking]
    #26389660 - 12/18/19 09:35 PM (4 years, 1 month ago)

I can send you a fresh print if you want as well :cheers: get everything back up and working


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OfflineTheApprentice
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Re: Q tips to transfer spores to agar. No germination [Re: cronicr]
    #26390626 - 12/19/19 12:13 PM (4 years, 1 month ago)

Quote:

cronicr said:
Get new spores noob





But they’re your KSSS bro bro :rofl:


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Invisiblealaskappalachian
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Re: Q tips to transfer spores to agar. No germination [Re: TheApprentice]
    #26390640 - 12/19/19 12:20 PM (4 years, 1 month ago)

I'd drop some in h20 and let the solution sit for a couple days- then swab the plate with the solution.  Older spores seem to like rehydration, ime.


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OfflinecronicrFacebook
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Re: Q tips to transfer spores to agar. No germination [Re: TheApprentice]
    #26390655 - 12/19/19 12:29 PM (4 years, 1 month ago)

Quote:

TheApprentice said:
Quote:

cronicr said:
Get new spores noob





But they’re your KSSS bro bro :rofl:



Oh shit:lol:


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It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor


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OfflineEdmunter
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Re: Q tips to transfer spores to agar. No germination [Re: cronicr]
    #26424897 - 01/09/20 03:06 PM (4 years, 20 days ago)

Quote:

cronicr said:
Quote:

TheApprentice said:
Quote:

cronicr said:
Get new spores noob





But they’re your KSSS bro bro :rofl:



Oh shit:lol:



No school like the old school......  #mic drop!


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