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OfflineRollo_grows
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Agar Spores don't germinate
    #27011432 - 10/30/20 03:40 AM (3 years, 3 months ago)

Hey everyone! I am a first timer using the Agar Tek and now some of my plates won't show mycelium growth...I hope you guys could give me some help :smile:
So I poured 10 plates with MYA Agar, waited for 1 hour to let them cool down, then inoculated them with a spore syringe and now one week after the inoculation there are still 5 plates where the spores won't germinate. The other half shows spots of mycelium growth, so the spores seem to be fine. What can I do now to fix it, should I just wait? I haven't used an inoculation loop, but will do it for the next time. I made sure to pure the Agar at 47Β°C temperature to avoid too much condensation.




I noticed when I used the spore syringe, that there is a big black clump of spores. I tried to shake it very hard, but they won't seperate. What do you usually do in this case, or doens't it matter? I continued anyways, hoping that it would work out.



Should I maybe inoculate them again? I would really appreciate your help :smile:


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OfflineSingularFusion
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Re: Agar Spores don't germinate [Re: Rollo_grows]
    #27011449 - 10/30/20 04:01 AM (3 years, 3 months ago)

i dunno why poor germination on some of your plates, maybe the clumping, but there is literally millions of spores in there if you can visibly see them

but i feel like it doesn't matter

what I mean is if half your plates germ well, the idea of agar in general will be to take the best couple of plates and make transfers to new plates. You will not need all of your germination plates

maybe one or two of the best ones to take your transfer material from

inoculating them again is not a thing IMHO, but you can of course noc some fresh plates instead if you feel like you need ten germ plates regardless

otherwise let them grow out a bit, get transferring and have some nice mush in short time

all the best


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OfflineCaptainParanoid
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Re: Agar Spores don't germinate [Re: Rollo_grows]
    #27011452 - 10/30/20 04:05 AM (3 years, 3 months ago)

I’m green so keep that in mind. If I’m wrong someone with more experience please correct me.

I’ve had this problem with not being able to break up the spores no matter how hard and long I shook that syringe. Searched for an answer and found that I needed an air bubble in there. My syringes had none.

The air needs to be sterile. The air needs to be drawn into the syringe while in a flame. I put the tip of the needle in the flame of my torch and drew the plunger back as much as I could (there wasn’t much room). Put the cap back on and with a bubble inside the clumps broke up easily. If you notice some of the water shooting out even with the cap on, don’t be alarmed.


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OfflineRollo_grows
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Re: Agar Spores don't germinate [Re: SingularFusion]
    #27018629 - 11/03/20 02:55 AM (3 years, 3 months ago)

Thanks a lot Nef, you're post and some more reading actually helped me a lot to understand the whole point of doing agar and isolating!!!
I used to think that I'll transfer the germinated plates directly to grain instead of doing some transfers to agar first. So this opened up a new world for me, thanks 😁

And also I believe my agar mix was too stiff, I used about 3% of agar, so maybe this was the reason. But actually it doesn't matter anymore. Will do some transfers today or tomorrow!


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OfflineRollo_grows
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Re: Agar Spores don't germinate [Re: CaptainParanoid]
    #27018631 - 11/03/20 02:57 AM (3 years, 3 months ago)

Thanks CaptainParanoid!
For fun purposes I'll try this the next time when using the syringe. But for now I believe doing transfers is the way to go for me :smile:
Have you actually tried to get in an air bubble and break up the spores? Also did you find any contaminations in your plates after doing that or was it a safe method?


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OfflineCaptainParanoid
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Re: Agar Spores don't germinate [Re: Rollo_grows]
    #27022089 - 11/04/20 06:42 PM (3 years, 3 months ago)

My syringes (4) were totally clumped up. When I tried putting them to agar I ended up getting big blobs of spores. Yes, I actually used the method above to get an air bubble in the syringe. It worked very well. The next time I used the syringe on agar there wasn't any contamination. Keep in mind the needle was red hot and I was using a Mapp gas torch not a tiny lighter. The needle tip was in the center of a very hot flame.


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