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Reviving 5yo Spore Print With Agar
    #29621816 -

Hi everyone, completely new to agar. Trying to revive nearly 5-year-old spore prints. Did a dry scrape across a few plates with no hydration; only this one single colony has popped up. It is currently 11 days since inoculation. How does the health look?

My Goal: Just want to get this to fruit so I can print fresh spores and salvage the genetics.

I have 3 pre-made plates right now (Full Nutrient (MEA)), and a larger batch arriving in about a week. (Also planning some LC later just for practice, but prioritizing this colony right now).

Questions:

  1.  Can I wait a week for my new plates? Will this colony be safe for another 7 days so I can do a bigger round of transfers, or should I use my 3 current plates immediately and do some transfers?

  2.  Go straight to grain? Should I skip a cleanup transfer and go straight to grain, or is a dry scrape from a 5yo print too high-risk for contamination?

  3.  Un-germinated spores: The mycelium is growing over areas with un-germinated spores. Will letting it run over them cause issues or drag dirty genetics/contam into my next transfer?

  4.  Timing & Selection: How far out should I let a single colony like this grow before it's ready to transfer? At what point does it become too early to work with, and at what point does waiting become too risky?




Extras: Filter Print Post Top
Re: Reviving 5yo Spore Print With Agar [Re: Natas420] * 1
    #29621831 -

Hi Natas :wave:

Great choice learning agar, and that growth looks pretty organized so my bet is that it's the right mycelium and not a contamination :hatsoff:

1: You can wait, but I like to do transfers from T0 plates early.

2: I would almost never go from T0 to grains, so I suggest you don't.

3: Yes is could, and why I do early transfers from T0 plates.

4: It's to early when you can't take a transfer with your scalpel, to late is a guessing game and always has a risk, like mentioned above transfer early.

So yeah there you have it, well my opinion on the matter anyway :muppet:


--------------------
“All Fungi are edible.

Some fungi are only edible once.”
― Terry Pratchett


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Re: Reviving 5yo Spore Print With Agar [Re: Kloakk]
    #29622010 -

Dude. I have heard that a couple hydrating the spores works wonder for old prints. But haven't tried it. I wish I had taken more prints as the prints I I have from 10 years ago don't seem to germinate and places like mushrooms  dot Com  went out of business. As a side note I think I'm personally responsible for the owner of mushrooms dot Com closing up shop.

Where the heck is a new good place to get spores.

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Re: Reviving 5yo Spore Print With Agar [Re: etuHad4] * 1
    #29622060 -

etuHad4 said:
Dude. I have heard that a couple hydrating the spores works wonder for old prints. But haven't tried it. I wish I had taken more prints as the prints I I have from 10 years ago don't seem to germinate and places like mushrooms  dot Com  went out of business. As a side note I think I'm personally responsible for the owner of mushrooms dot Com closing up shop.

Where the heck is a new good place to get spores.


The marketplace sub forum right here on the org :awesomenod:


--------------------
“All Fungi are edible.

Some fungi are only edible once.”
― Terry Pratchett


Extras: Filter Print Post Top
Re: Reviving 5yo Spore Print With Agar [Re: Kloakk]
    #29625487 -

Hello Kloakk,

I hope all the extra effort of learning agar is worth it! It's definitely been a lot more expensive and stressful than what I'm used to (MSS to BRF tek), but I'm enjoying the learning process.

I've done a several transfers now, and the mycelium was definitely healthy. I just wish I had a flow hood—I'm limited to working in a SAB at the moment.

What's the best method for going from agar to grain when you're only using a SAB?

My pressure cooker is quite small, so I can only fit quarter-pint jars. I can buy pre-sterilized grain bags, but I don't think I could fit one inside my SAB to inoculate it safely.

Another option I was considering is using pre-cooked rice, snipping the top open, adding a few agar wedges, and taping it back up. I could also make some quart jars with grain (I'm not sure which type would be best), pressure cook them myself, crack the lids open in the SAB, and drop in a few agar wedges. I also have plenty of BRF ingredients (I steam my BRF jars for sterilization). I have several syringes available too, so I was wondering whether making a Liquid Inoculant (LI) or mycelial slurry from the agar would work well for BRF jars. I'm also considering making some liquid culture (LC), if that might be a better option. Or is there another approach I haven't considered yet?

Thanks for your help—I really appreciate any advice!

Edited by Natas420 (07/16/26 07:18 AM)

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Re: Reviving 5yo Spore Print With Agar [Re: Natas420] * 1
    #29625534 -

Hi Natas :wave:

Agar is definitely worth it, it feels awkward at first, but you'll get used to it. I suggest doing one wedge per jar when you're working in a SAB, that way you can have your jars sitting on the "floor" of the SAB behind the rack and the plate on the rack like this.



I know this is a FFU, but I did the exact same thing in the SAB except with the FFU I work behind the jars and not from the side.

I'll wipe down the lids, unscrew it half a turn just so I don't have to use force inside the SAB, then I'll put my jars inside the SAB, wipe the plate and place inside SAB. Unscrew the lids so they are resting on the jar, unwrap plate and put it upside down. Lift the bottom of the plate and cut all the wedges, I do it like this.



After making the cuts I take the first wedge on my scalpel and put the plate back down on it's lid, still upside down. Then tilt the lid enough to easily get the wedge inside the jar and rest the lid on the jar again. From there you could either flame the scalpel or go for another wedge, until you feel comfortable with the movement and don't have everything invested in your jars I suggest flaming between wedges, but your call.

After all wedges are in all 4 jars I turn the plate the right way up and then close the jar lids, but if you're going to flame the scalpel between jars, close each lid after the wedge is in. Have a sucky gif on how I work with plates, might help :muppet:



--------------------
“All Fungi are edible.

Some fungi are only edible once.”
― Terry Pratchett


Edited by Kloakk (07/16/26 10:34 AM)

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Re: Reviving 5yo Spore Print With Agar [Re: Kloakk]
    #29650581 -

Hey Kloakk!

Thanks for your help! I followed what you suggested, I sent one plate per jar(hopefully not too much but had available clean plates) and this is the result. It’s been about 8–9 days since the agar-to-grain transfer. The jars I’m using were pre-sterilized millet with 1.9-micron GE filter holes.

The plates had quite a lot of aerial mycelium, but smelled strongly of mushrooms, so I decided to transfer the mycelium to the grain. How does it look in your opinion, and should I do a break and shake at this point?

I currently have the jars sitting in a 50L tote. I’m not sure if that’s the best way to store them, or if they need much fresh air at this stage. I assumed they wouldn’t need much, since higher CO₂ is preferred before fruiting, but please correct me if I’m wrong. I can always just keep them on a shelf in a cupboard or something instead.

Thanks again for all your help — I really appreciate it!






Edited by Natas420 (08/18/26 09:52 PM)

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Re: Reviving 5yo Spore Print With Agar [Re: Natas420]
    #29650605 -

Morning Natas :goodmorning:

Jars look good from here and yes you can do a break&shake :rockon:

I would try to move the jars around slowly at first and get as much millet stuck to the agar as possible before you do the shake to increase the chances of not getting the agar stuck to the jar or lid.


--------------------
“All Fungi are edible.

Some fungi are only edible once.”
― Terry Pratchett


Extras: Filter Print Post Top
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