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Invisiblec10h12n2o
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Forced hydration of spores
    #26492778 - 02/19/20 03:00 AM (3 years, 11 months ago)

Hi everyone. So I'm trying to get get some old spores to germinate, never had any luck with them on agar

I figured I would try the forced hydration method with syringes and rubber bands.

So i sterilized some distilled water and a few drops of jet dry, scraped some spores into a petri dish, and put about 1ml of water onto it before suckimg it back into the syringe. Then I capped the syringes and put rubber bands on them, as per stros tek

After about 9 hours I did the calculation and realized that for the 3ml syringes I am using,  I put the rubber bands on waaay too tight, since only 1lb of pressure on the plunger already generates over 10psi.. I had it super tight. I now only have one less thick rubber band on each syringe

Hopefully I didnt cause cytolysis in the spores :/

Does anyone have any experience with this (or another) method working to revive some old spores? If so, how long should I leave them under pressure before I put a drop on agar?

Thanks, really hope I can germinate these things


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InvisibleHobbyist
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Re: Forced hydration of spores [Re: c10h12n2o]
    #26492818 - 02/19/20 04:45 AM (3 years, 11 months ago)

Dude, it's super good to see you back! :laugh:

I've read about a guy mentioning that he suspended the spores in sterile water for 6-8 months and then he was able to get some viable spores to germinate.  It was somewhat recent but I can't for the life of me remember who posted it.  They referenced stro's method in their post so you could probably pull it up with some fancy use of the search engine.


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OfflineSynKyd
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Re: Forced hydration of spores [Re: Hobbyist]
    #26493051 - 02/19/20 08:56 AM (3 years, 11 months ago)

I have a similar situation. My plan is to let them soak a few weeks in the syringe then try some plates. If I get no growth I will start with light pressure for a week and try putting to agar again, increasing pressure slightly each week until something happens.

Seeded my syringes around 2/9 so we shall see soon, soon as I make up some fresh plates!

:hippie:


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OfflineKizzle
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Re: Forced hydration of spores [Re: SynKyd]
    #26493194 - 02/19/20 10:44 AM (3 years, 11 months ago)

I'm not sure there's any merit to that method. You might try different agar recipes. Things like pH can affect whether a viable spore germinates or not and individual spores can vary in exactly which conditions will activate them.


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OfflineSyntax.Error
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Re: Forced hydration of spores [Re: Kizzle]
    #26493308 - 02/19/20 12:01 PM (3 years, 11 months ago)

Dont mean to Hijack but its kind of on topic.
In years what would you guys say is "old" for spores?


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OfflineSynKyd
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Re: Forced hydration of spores [Re: Kizzle]
    #26493341 - 02/19/20 12:18 PM (3 years, 11 months ago)

Quote:

Kizzle said:
I'm not sure there's any merit to that method. You might try different agar recipes. Things like pH can affect whether a viable spore germinates or not and individual spores can vary in exactly which conditions will activate them.




That’s a great point on the agar, and is also part of my plan.

My spores are 5 years old and doing a basic print scrape to agar has given only 2 potentially viable starts with probably 15-20 misfires.


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Invisiblec10h12n2o
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Re: Forced hydration of spores [Re: Kizzle]
    #26493353 - 02/19/20 12:28 PM (3 years, 11 months ago)

Quote:

Hobbyist said:
Dude, it's super good to see you back! :laugh:

I've read about a guy mentioning that he suspended the spores in sterile water for 6-8 months and then he was able to get some viable spores to germinate.  It was somewhat recent but I can't for the life of me remember who posted it.  They referenced stro's method in their post so you could probably pull it up with some fancy use of the search engine.




Thanks buddy :smile:
Quote:

Syntax.Error said:
Dont mean to Hijack but its kind of on topic.
In years what would you guys say is "old" for spores?




Some are as young as 4 , others around 8yrs

I've germinated lots of other stuff in that range without issue, but these are giving me lots of trouble. Shoulda probably just hydrated before trying forced hydration,  but I'm impatient


Quote:

Kizzle said:
I'm not sure there's any merit to that method. You might try different agar recipes. Things like pH can affect whether a viable spore germinates or not and individual spores can vary in exactly which conditions will activate them.




Thanks for the reply. I actually did try another recipe before , and played with the ph (not much but a little), and tried activated charcoal, all with no luck.

This time around I just streaked them again and with no luck after 15 days I figured I'd try hydrating them

What makes you think the forced hydration method has no merit? Seemed to work well for RR, stro, and mycolorado


--------------------

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche


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Offlinespiritlands
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Re: Forced hydration of spores [Re: Kizzle]
    #26493383 - 02/19/20 12:51 PM (3 years, 11 months ago)

Quote:

Kizzle said:
I'm not sure there's any merit to that method. You might try different agar recipes. Things like pH can affect whether a viable spore germinates or not and individual spores can vary in exactly which conditions will activate them.




Ya hydration didn't work for me at all. The longest I hydrated for was over a month. The only thing that was working for me was liberal use of spores and transferring any colonies that appear. Most of the prints were from 2012. For me anything that was going to germinate did so within three weeks. I waited on pitris for close to two months with no growth.

This was an x7x print from 2012 and you can see how much spores it took over several pitris. The print is about 3in in diameter. The cube prints seem most resilient but I have had several pan prints that were dead and some other exotics.


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OfflineSockadin
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Re: Forced hydration of spores [Re: c10h12n2o]
    #26493410 - 02/19/20 01:16 PM (3 years, 11 months ago)

What about starting with an alkaline water, no I'm not one of those water Nazis people who drinks ionated water. Just curious what the optimum PH would be?


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InvisibleInfiniteDreams
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Re: Forced hydration of spores [Re: Sockadin] * 2
    #26493658 - 02/19/20 04:11 PM (3 years, 11 months ago)

Waterboarding, it's what spores crave


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Invisiblec10h12n2o
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Re: Forced hydration of spores [Re: InfiniteDreams]
    #26493750 - 02/19/20 05:17 PM (3 years, 11 months ago)

if anyone knows an optimal ph for germination i can try that again, i have the equipment to do it right.. just had no luck with what i tried before


--------------------

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche


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OfflineShabbyRabbit
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Re: Forced hydration of spores [Re: c10h12n2o]
    #26932213 - 09/12/20 05:23 PM (3 years, 4 months ago)

Did ya have any luck here?

I'm in a similar spot and looking for the best tek at the moment.  :b00ftube:


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