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Rehydrating old spores


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#1 stackedandeasy

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Posted 05 April 2021 - 07:04 PM

I have a whole bunch of prints of pan that are a few years old that I want to rehydrate, any suggestions. I've already streaked a dish with some but they seemed pretty dry and I thought I'd try to hydrate some too. Thanks for any help.



#2 Auhron

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Posted 05 April 2021 - 07:11 PM

Make yourself a spore syringe, let it rest a day or two, then use it as usual.  I recently germinated 9.5yr old spores without doing anything other than putting them to agar. They just took a long time to show signs of life. If the spores are rehydrated first germination should be a fair deal faster.

 

Ensure you use filtered, sterilized water. Good luck!


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#3 Sidestreet

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Posted 06 April 2021 - 04:26 AM

I'm rehydrating 4-year-old Cubensis spores in filtered, sterilized water.  I'm planning on giving them a week and then doing a single PF jar for each variety to test for viability and cleanliness.  It's the first time I've attempted it, so fingers crossed.


Edited by Sidestreet, 06 April 2021 - 04:26 AM.

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#4 stackedandeasy

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Posted 06 April 2021 - 01:33 PM

Sterilize water then scrape some spores into it and draw them back into a syringe then inoculate the agar?  I thought about just adding a few drops of sterile water to the print and cover it for 24 hours, would that not work?



#5 stackedandeasy

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Posted 06 April 2021 - 03:04 PM

I also had two liquid cultures that has been sitting for three years, I thought the bigger one might have a bacteria but now I'm not sure, I pulled from both of them and inoculated a couple of dishes with the browner one, I'm thinking of inoculating a few of the dishes with the lighter one today. I just sterilized some water to make a syringe.  

 

 IMG_2960.jpg IMG_2958.jpg IMG_2959.jpg



#6 stackedandeasy

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Posted 06 April 2021 - 04:46 PM

The dishes I incolulated 24 hours ago with the liquid culture with all the mycelium floating in it. In case you're wondering why the culture is brown is because I incoluted it with agar made with manure.

 IMG_2966.jpg IMG_2962.jpg IMG_2961.jpg



#7 Sidestreet

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Posted 06 April 2021 - 05:39 PM

Sterilize water then scrape some spores into it and draw them back into a syringe then inoculate the agar?  I thought about just adding a few drops of sterile water to the print and cover it for 24 hours, would that not work?

I PCd some water in a pint jar with a SHIP lid on it, and also a syringe.  Under sterile conditions, I drew a syringe full of water and then shot it into the button bag containing the spore print.  I agitated the button bag with the print and water until the water was purple.  Then I drew it back into the syringe and shot that back into the pint jar with the rest of the water.

 

I suppose I could have shot the water into the spore print bag and let it sit there for 24 hours, but this way it's done all at once and I have a bunch of spore water ready to go.

 

Good luck with your liquid cultures!


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#8 stackedandeasy

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Posted 08 April 2021 - 03:05 PM

Yeah my liquid culture isn't contaminated! I just started a batch of rye soaking and I'm going to inoculate it with the liquid culture plus I'm going to make a new one using new agar plates. The spores haven't started yet but it's only been a few days. I'll keep my you updated on how it's all coming up, geeze I love fresh pan for the summer!IMG_2988 (3).jpg IMG_2989 (2).jpg


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#9 stackedandeasy

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Posted 09 April 2021 - 08:27 PM

Dishes look awesome for just a few days old, I started cutting a few wedges out of one of them and inoculated another dish with the other liquid culture to see if it's good, I thought it may have a bacteria because the water looked cloudy but it settled and looked pretty clear so we shall see. I sterilized a few jars of rye and inoculated them with the liquid culture. I may start a new thread with pan grow as the topic.

 IMG_2996.jpg IMG_2997.jpg


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#10 stackedandeasy

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Posted 10 April 2021 - 06:45 PM

One of the dishes with the spores is showing signs of life, they were at least 5 years old, some were from 2011. IMG_3013.jpg


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