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OfflineJunior Fungus
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Rehydrating a Crispy Old Plate
    #23854713 - 11/21/16 03:46 PM (7 years, 2 months ago)

Gonna start off by saying I love you guys. :heart:

I had this plate that I left in the fridge for a year. Great genetics on it. I want to save it.

Here's my plan:
1. Soak it in a 1:10 sterile bleach solution for an hour, swirling it.
EDIT: The bleach step should have been a few minutes, not an hour, and may not have been necessary. It is only appropriate if contams are visible. Mycelium can survive this treatment, but contams don't. However, the mycelium is weakened and takes a week or two to recover from the bleach. Reference: RR, https://www.shroomery.org/forums/showflat.php/Cat/0/Number/5599236
2. Transfer it to a fresh jar of sterile water, soak for a certain amount of time (till it looks nice and plump I guess).
3. Transfer pieces to a dozen plates, clean up the cultures if mycelium is visible.
4. SLANTS! so that I don't have to do this ever again.
5. Inoculate some WBS!

Am I missing anything here? Any tips for this kind of procedure? Please be specific.


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Edited by Junior Fungus (11/22/16 08:28 PM)


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OfflineKenetic
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Re: Rehydrating a Crispy Old Plate [Re: Junior Fungus]
    #23854783 - 11/21/16 04:05 PM (7 years, 2 months ago)

bleach would probably kill a weak, dehydrated culture.  What else can I say?


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OfflineJunior Fungus
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Re: Rehydrating a Crispy Old Plate [Re: Kenetic]
    #23854825 - 11/21/16 04:18 PM (7 years, 2 months ago)

I picture the bleach only killing the contaminants on the outside of the mycelium. Do you think that's impossible or very unlikely?

I mean there was no water in this plate at all, so I figure the bleach wasn't there long enough to penetrate and kill all the mycelium. All I need is for the contams to die off, and for a bit of mycelium to survive.


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InvisibleJosex
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Re: Rehydrating a Crispy Old Plate [Re: Junior Fungus]
    #23854847 - 11/21/16 04:24 PM (7 years, 2 months ago)

Love you too :heart:

If I were you I wouldn't use bleach at all. I'd start from the simplest procedure and go my way up from there if it fails.

You could start by simply transferring a piece of myc to another plate and then try the "sandwich tek" if transferring normally fails.

It is possible to "resurrect" myc from a dried up fruit so I don't think you're going to have any issues with that plate.


Edited by Josex (11/21/16 04:25 PM)


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InvisiblePastywhyteMDiscord
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Re: Rehydrating a Crispy Old Plate [Re: Josex]
    #23854861 - 11/21/16 04:29 PM (7 years, 2 months ago)

I would drop the plate in a jar of sterile water to rehydrate. The agar will also release the nutes into the water as it hydrates allowing any potentially live myc to take off. Then aspirate and inoculate a fresh plate to determine viability.


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Re: Rehydrating a Crispy Old Plate [Re: Pastywhyte]
    #23854905 - 11/21/16 04:42 PM (7 years, 2 months ago)

Fuck it, good idea Pasty. I have a KSSS culture on a BRF agar plate, dried af and it's just sitting on a shelf, not even in the fridge. I'm going to try that.


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OfflineJunior Fungus
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Re: Rehydrating a Crispy Old Plate [Re: Pastywhyte]
    #23855062 - 11/21/16 05:40 PM (7 years, 2 months ago)

Quote:

Pastywhyte said:
I would drop the plate in a jar of sterile water to rehydrate. The agar will also release the nutes into the water as it hydrates allowing any potentially live myc to take off. Then aspirate and inoculate a fresh plate to determine viability.




I'll give that a shot before transferring pieces to fresh agar. Any idea how long I should let this thing soak?

I did the bleach thing before even making this thread. Now it's been soaking for a few hours in sterile water, and its looking nice and hydrated already... especially the in vitro pins.


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InvisiblePastywhyteMDiscord
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Re: Rehydrating a Crispy Old Plate [Re: Junior Fungus]
    #23855123 - 11/21/16 06:02 PM (7 years, 2 months ago)

Why bleach at all? It was a clean plate, all bleach is going to do is hurt. In fact why make the thread in the first place if you already soaked it? Confusing.


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OfflineJunior Fungus
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Re: Rehydrating a Crispy Old Plate [Re: Pastywhyte]
    #23855875 - 11/21/16 10:31 PM (7 years, 2 months ago)

My train of thought was that as the plate dried, it weakened the mycelium and let contams develop. I thought I saw small signs of contams, so I bleached.

In hindsight, I might have misdiagnosed.  So I should have just skipped the bleach, I think you're right. I thought of making this thread after starting the procedure. I just wanted to see new opinions on rehydrating, as I had used the search function a lot already.


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InvisiblePastywhyteMDiscord
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Re: Rehydrating a Crispy Old Plate [Re: Junior Fungus]
    #23855888 - 11/21/16 10:39 PM (7 years, 2 months ago)

Why would contams develop if it was never exposed to them? Contam vectors are specific and come from somewhere. Contams do not just form out of nothing.

It's your grow to do with what you wish. But I wouldn't hold my breath for this particular one. If it works great. But if not then get busy.


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OfflineJunior Fungus
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Re: Rehydrating a Crispy Old Plate [Re: Junior Fungus]
    #23898475 - 12/05/16 10:12 PM (7 years, 1 month ago)

UPDATE: Success!

The plate that got bleached has produced some growth after a transfer. I just transferred the growth to a fresh plate. It might not actually be mycelium... it's quite fuzzy. Oh and the original plate wedges started to turn brown, and the pins too. It all looks like it's rotting.

I took Pasty's advice for another crispy old plate I had and just soaked it in sterile water directly. I soaked it overnight and then transferred wedges to fresh plates. Within 5 days, I saw nice, clean rhizomorphic growth!

In conclusion, the bleach killed most of the first plate I tried reviving, so I would advise against using bleach for this procedure. Simply rehydrating a plate in sterile water was enough to get clean growth.

Thanks Pasty :thumbup:


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