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Offlinerhapsodyclaymore
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New grower, longtime lurker - starting with agar with an old print - have some questions
    #27281014 - 04/25/21 04:15 AM (1 month, 18 days ago)

Hi Shroomery community. Longtime lurker, casually, but a few weeks ago decided to finally grow.

Happy to be here. Let me explain what I have. I've got a 3.5 year old GT print I hand picked up at a shop (reputable dealer) that I had wanted to use for growing. It just needed a long time to decide to get the supplies and begin (started from scratch). I've got another print that's likely 25 years old which was hand given to me by an old supplier of shrooms back in the late 90s and is his own custom hybrid strain. For ease, I've decided to not end in disappointment have focused on the GT print I have first. Both prints were stored in a shoe box out of light and heat sources.

After reading for weeks, I decided the best way forward was to do agar. 5 weeks ago, I tried this method:
potato flakes: 12g
agar agar: 8g
corn syrup: 4g
2 cups water

Simmered on the stove, poured into 2x 250ml media bottles and PC'ed at 13psi for about 35 minutes. Used a laser temperature reader to wait for it to cool down to around 54C before pouring using oven tek into 2oz condiment cups (25 total) and then stacked to avoid condensation buildup.

Next day in a SAB scrapped about 25% of a spore print into 12 of the cups. One of the 13 unused cups within 2 days got contaminated. Presently after 5 weeks now, all the used and unused cups have no contams.

The only cups of the 12 are 2 that have something going on.

This one here started up after around 4 days and while these images are now 1.5 weeks old (3.5 weeks from spores added), it has grown out roughly 20% more, but hasn't yet overtaken the cup.



This one here started out like this at 3.5 weeks old (taken 1.5 weeks ago)


But now since last week, from one day to the next day, it turned white:




That was my first attempt. Then I did some more reading and I do realize that the print is a few years old and the spores are a bit dry, but probably still good I think.

My next experiment with agar:
Then I was suggested to try no pour agar with less agar to keep more liquid inside which could help the spores hydrate. I purchased 28x 53ml glass canning jars and used:

500ml distilled water
10 grams Light Malt Extract
5 grams agar agar powder

Brought to a simmer and poured into the jars. Tops put on with half screw on to allow air flow, and individually wrapped with foil on top to prevent water getting in, and then PC'ed for 35 minutes before letting cool down for a few hours before opening and screwing the tops on. After 4 days, no contam in any of the jars.

Yesterday I took 12 of the jars and added about 25% of the GT spore print to them in my SAB. There's nothing really to show yet, but I hope in a few days/weeks something will appear.

My plan for both of these creations will be to add to 250ml canning jars with whole oats  to spawn, and then put to bulk substrate in a monotub. That's the plan right now.

As a question to the community, getting the agar out of the glass jars could be a problem. I plan on cutting the agar up with a hobby knife (flame sterile) into square chunks, but I don't think they will easily fall out of the jars. Is it acceptable to use a sterile small spoon to scoop out the parts into the spawn jars when the time is right? Or what is the recommended way?

Whatever happens at the end, I will be making new spore prints from the biggest caps that I can get to grow from both of the prints.

Is this a good path? I have seen numerous threads here on old prints and results do vary, and they do take a long time to happen, but I'm patient. Any suggestions or comments are always welcome. Thanks for taking the time to read over a new grower's work. I'm just looking to have 1 way that works which is repeatable each time that I can rely on.


Edited by rhapsodyclaymore (04/25/21 09:48 AM)


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27283333 - 04/27/21 02:48 AM (1 month, 16 days ago)

Anyone have any thoughts on the growth in my cups?


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InvisibleGoatriderS
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27283369 - 04/27/21 04:09 AM (1 month, 16 days ago)

What i can see is slimy bacterial bubbles or something?
These embossed lids make it hard to judge, also no good quality.

If i had old spores, i`d make up a syringe for hydrating them,
and some brf pucks.
With shooting the spores on brf, you`ll have a great chance to get some myc,
you can transfer to agar then for further cleanup.

Read here for the Josex Poke, i really recommend it, if you want to save your cultures.

                    :cookiemonster:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Goatrider]
    #27283418 - 04/27/21 06:43 AM (1 month, 16 days ago)

Wonderful suggestion! I thought to make a syringe, but then tried a different agar formula with a more liquidy with the hope that the spores hydrate better. Thanks for the tip!

What I'll do is wait a bit and see if the new jars I made do something. If I don't see anything in 2 weeks, in parallel I'll work on a spore syringe. But, I'll need to get some syringes.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27287447 - 04/30/21 05:52 AM (1 month, 13 days ago)

7 days later after spores were added to the new agar batch (colored red to differentiate from the blue 2oz cups), I woke up to find that 1 of the 12 no pour jars had fuzzed up a little.

I cannot get a cleaner pic than this. Tried for 10 minutes.

Progress?



In case these spores don't work in the end, I've ordered a new GT print as a backup to use.


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InvisibleGoatriderS
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27287457 - 04/30/21 06:33 AM (1 month, 13 days ago)

Why that thick agar?
That would have been enough for about 10 plates :smirk:

Wait a few days more, 7 days isn`t much time for spores on agar.
Post more pictures next week.

            :cookiemonster:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Goatrider]
    #27287494 - 04/30/21 08:01 AM (1 month, 13 days ago)

So, I made too much and then decided to top off some of the containers. Looking back at that, it was a bad decision. Also I ran out of jars so it wasn't like I could just keep pouring. Made 28 jars.

To... properly get it for moving to a spawn jar, I can slice it up (tic-tac-toe style) and if it won't come out, is it possible to just use a sterile small spoon to scoop it out?

Pardon the noob questions. I'm learning :smile:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27288584 - 05/01/21 03:56 AM (1 month, 12 days ago)

Short update from overnight. In the jar, there's now 25x more of what I photographed on the surface. Nothing yet from the other jars.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27291696 - 05/03/21 11:18 AM (1 month, 10 days ago)

Hey everyone,

There was a mixup at the shop and I was mailed a 12ml GT spore syringe instead. I've not done much research with syringes and agar plates. I've read just a tiny drop just poking under the surface of the center of the agar is fine, ensuring that it doesn't roll around. I've also read that streaking is good too. What is the best way for me to easily use my remaining agar with the syringe?

The syringe is currently sitting in the refrigerator until I'm ready to use it.


Edited by rhapsodyclaymore (05/03/21 11:20 AM)


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OfflineFungi Rogers
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27292232 - 05/03/21 04:56 PM (1 month, 10 days ago)

I put a drop on my inoculation loop and then streaked my plate with it. Worked pretty good. You could also just put a single drop on it and let it do it’s thing.


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OfflineBig_Dub
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Fungi Rogers]
    #27292450 - 05/03/21 07:01 PM (1 month, 9 days ago)

Yeah just put a single drop.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Big_Dub]
    #27293341 - 05/04/21 06:44 AM (1 month, 9 days ago)

Great! Just wanted to confirm!

I've got 12 of the blue agar plates left (flipped upside down to keep the moisture inside) without contam. They are around 6 weeks old now. And I've got 18 of the red no pour agar jars in reserve too. I want to do maybe 4 of each with the syringe and see how they go, leaving the others in reserve should contam happen, then I can transfer away with the remaining agar plates that I have.

I'm still not giving up hope that the other agar plates will show signs of life. It's a good experiment to see if I can successfully resurrect an old print. I really want to try this, if successful this time, with the double decade old one I have.

Thanks for the feedback. I'll get this eventually!


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OfflineFungi Rogers
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27293364 - 05/04/21 07:32 AM (1 month, 9 days ago)

Making softer agar will sometimes help revive old prints. But if you take the old print and make a syringe out of it. It should revive it as best as it can.

What I like to do when coming from spores is put 3 transfer to a new plate spaced out evenly that way instead of using 3 plates you use one.  Then just find the growth your liking.


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InvisibleGoatriderS
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Fungi Rogers]
    #27293385 - 05/04/21 07:43 AM (1 month, 9 days ago)

https://www.shroomery.org/forums/showflat.php/Number/25162436

I tried it once, and it worked perfectly.
If nothing helps, you may try this.

            :cookiemonster:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Goatrider]
    #27294174 - 05/04/21 05:08 PM (1 month, 9 days ago)

Ah, so basically 3 tiny drops in a circle, and then pick the best source and migrate to a new plate? Sounds like a good plan. Good tip! I'll employ that on my Saturday in my SAB and hope for the best in a few days. I'm feeling pretty good that this batch will work out and my older plates (with the old spores) could come alive... maybe... in a few days/weeks. I'm open to keeping the older ones active like that just to see how it goes. For science!


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OfflineFungi Rogers
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27294393 - 05/04/21 07:33 PM (1 month, 8 days ago)

Quote:

rhapsodyclaymore said:
Ah, so basically 3 tiny drops in a circle, and then pick the best source and migrate to a new plate? Sounds like a good plan. Good tip! I'll employ that on my Saturday in my SAB and hope for the best in a few days. I'm feeling pretty good that this batch will work out and my older plates (with the old spores) could come alive... maybe... in a few days/weeks. I'm open to keeping the older ones active like that just to see how it goes. For science!





I mean you could do it with drops I was more referring to when you’re transfer mycelium from the germ plate to a new plate.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Fungi Rogers]
    #27295304 - 05/05/21 11:39 AM (1 month, 8 days ago)

Quote:

Fungi Rogers said:
I mean you could do it with drops I was more referring to when you’re transfer mycelium from the germ plate to a new plate.




OH! This actually makes more sense. Thank you for the clarification.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27299931 - 05/08/21 11:27 AM (1 month, 5 days ago)

Alright. So, getting a drop out of a syringe is harder than I thought. Kept getting stuck a bit inside the needle. For 8 cups (4 blue, 4 red), I've used about 1.8cc from the syringe. Let's see how the next days go. I may have used more than 1 drop in each one accidentally.

Anyway, this is my first time. It's a learning process.


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InvisibleGoatriderS
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27299953 - 05/08/21 11:41 AM (1 month, 5 days ago)

Try to squeeze the barrel of the syringe to force a drop out,
instead of pushing the plunger.
I always do this, if i have to use a syringe to agar.

            :cookiemonster:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Goatrider] * 1
    #27300029 - 05/08/21 12:55 PM (1 month, 5 days ago)

Quote:

Goatrider said:
Try to squeeze the barrel of the syringe to force a drop out,
instead of pushing the plunger.
I always do this, if i have to use a syringe to agar.

            :cookiemonster:




Oh that's genius! I never even considered that. The next time I need to use the syringe, I'll be sure to do it that way. Thank you!


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27301032 - 05/09/21 05:00 AM (1 month, 4 days ago)

Update today: in the 4x4 (blue/red) agar cups I've used the spore syringe with, there are now between 1-4 tiny white dots on the surface of each cup. It was mild yesterday, barely there, but now today they are more pronounced. One of the blue agar cups has what appears to be a white "S"-shape on the surface. I'm not sure if this is contam or not or some sort of bacterial infection.


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InvisibleGoatriderS
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27301069 - 05/09/21 06:09 AM (1 month, 4 days ago)

If you inoculated yesterday, it`s not usual to see mycelium growth
24 hours later. Or was it earlier?
Don`t wanna say it won`t happen, but watch them carefully.
Feel free to load up some pics.

You didn`t streak out with a loop or swab, as you say s-shape?
Sounds like bacteria, but that doesn`t matter at that stage.
Inoculation plates are always there to clean up and transfer from.
Just give pictures :wink:

        :cookiemonster:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Goatrider]
    #27301169 - 05/09/21 08:37 AM (1 month, 4 days ago)

Actually, in maybe 2-3 hours it will be almost exactly 24 hours.

The "S" thing is really small, tubular in shape, and white. Could be bacteria. If I remember correctly, the blue agar plate I threw away from the initial 25 cups had that too. This is why I did 4x4 of the plates (red/blue) from what I did so that I can transfer out the good stuff into fresh plates.

I'll see perhaps tomorrow to upload some pics should there be something more amazing happening.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27302365 - 05/10/21 04:34 AM (1 month, 3 days ago)

OK, photos.

GT Spore syringe, same agar plates used. around 45 hours after inoc.





The blue ones are more solid with double the agar used compared to the red ones I was trying a different formula on. You can see bumps on the surface of the red ones which are more under the surface than the blue ones where the myc is on the surface. Either way, good growth so far.

I've put aside the older ones I did weeks ago with the hope that they might come alive in a few weeks or months.


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OfflineSamushlove
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Goatrider]
    #27302714 - 05/10/21 11:24 AM (1 month, 3 days ago)

Good idea:thumbup: that can be challenging


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: Samushlove]
    #27303791 - 05/11/21 05:51 AM (1 month, 2 days ago)

Blue ones are still taking off still. Red ones are more below the surface of the agar since 50% agar was used. I assume that it doesn't really matter if the myc is visible as white fuzz on the top of the agar or not. Myc is still myc.

No contam signs yet, so far so good!


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27305470 - 05/12/21 11:28 AM (1 month, 1 day ago)

Small update: blues are going wild still. The red ones, given the more liquidy agar formula, took a while to fuzz through the surface, but 3 of 4 now have fuzzed either a little or a lot. Good sign.


I'll try to take some photos tomorrow to show the progress. I'm excited.


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27306632 - 05/13/21 07:43 AM (1 month, 9 hours ago)

Ok, here is the current state. Looking good, i think. I probably need to do some transfers to new agar (for more agar) this weekend, and maybe in another week or two I can move to spawn jars. I ordered some whole oats this week.





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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27308273 - 05/14/21 01:43 PM (30 days, 3 hours ago)

Growth is still going well. But, I have a question: I'd like to make a few transfers to clean new agar (just a few or so) as I'll need a few more to properly inoc my 12x 250ml jars i have for spawn. Can I leave the agar that I've removed sections from alone and let it continue to take over the agar and use in the spawn jars? Is my growth doing well, fine? Just looking for a little input on this. Happy no contam :smile:


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Offlinerhapsodyclaymore
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Re: New grower, longtime lurker - starting with agar with an old print - have some questions [Re: rhapsodyclaymore]
    #27309382 - 05/15/21 08:07 AM (29 days, 9 hours ago)

This one, of the agar cups, looks to be the best one for use to transfer to others. I picked 3 slices from the top of the cup and moved them today to 3 new blue cups.




After transfer. This was my first time. I got the hang of it better on the 2nd cuts.




This should hopefully take the best rizo....morphic(?) bits.


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