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OfflineDreadedBee
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Monokaryotic culture from MS syringe?
    #27148570 - 01/14/21 08:11 PM (3 years, 1 month ago)

Hi all,

Very long time lurker, just signed up so Hello and thank you to everyone who contributes, can't give enough praise for the wealth of knowledge available here.

I started a few agar dishes (small pp5 containers using only agar and grain water) 8 days ago from MS syringes sourced from a local vendor. 6 in total - 2x Tasmanian, 2x Ecuador and 2x B+.

I forgot to shake the B+ syringe. The Tasmanian and Ecuador which I did shake  have multiple germination sites.

One of the B+ plates has shown no signs of growth, the other has a single germination point which is growing very aggressively and very uniform, looks like a monoculture based on my limited experience. I'm considering the possibility that no spores reached the first plate, and only 1 reached the 2nd.

I've read the boards pretty thoroughly and understand that isolation of a single spore is highly unlikely based on my procedure, but as always there is lots of conflicting opinions. Based on the below images I'd love to hear all of your thoughts. Apologies for the image quality.



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OfflineTstone
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Re: Monokaryotic culture from MS syringe? [Re: DreadedBee]
    #27148613 - 01/14/21 08:43 PM (3 years, 1 month ago)

It's the only place that germinated, not a mono culture. You don't need 20 drops on a plate. Single drop, all that is needed. Spores are microscopic, even not shaking the syringe, spores are everywhere in the syringe.

With messy plates, like the others, your going have a tough time trying to decipher  nice growth for future transfers.  A single drop per plate, keep it flat so the spore solution isn't running all over your plates. The poor quality photos, hard to tell if you have mycelium. What's the black  on the edge of the B+ dish? Spores?


Edited by Tstone (01/14/21 08:45 PM)


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OfflineDreadedBee
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Re: Monokaryotic culture from MS syringe? [Re: Tstone]
    #27148628 - 01/14/21 08:50 PM (3 years, 1 month ago)

The marks on the B+ image are permanent marker and the injection molding point on the lid. There are no marks on the agar.

I have around 40 plates running at the moment from the same sab session. I'm fairly confident what is pictured is mycelium.

The growth of this B+ is far more uniform than my gt and bspk plates after 3 transfers. Is this just B+ genetics?


Edited by DreadedBee (01/14/21 09:05 PM)


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OfflineCyonic
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Re: Monokaryotic culture from MS syringe? [Re: DreadedBee]
    #27179619 - 01/31/21 04:02 PM (3 years, 16 days ago)

You have to view the hyphae and through a microscope to be able to tell if you have a monokaryon. If you do you can tell by the clamp connection of the hyphae cells.

I would not think that one could tell by looking at the growth of the mycelium on agar, other than that if you have very rhizomorphic growth it is likely dykariotic, and not monokariotic.

But even some dikaryiotic mycelium is not at all rhizomorphic.


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OfflineStkn_Hywd_Myco
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Registered: 08/31/20
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Re: Monokaryotic culture from MS syringe? [Re: Cyonic]
    #27187686 - 02/05/21 09:20 AM (3 years, 11 days ago)

Make some transfers and find out, but it's highly unlikely that its a monoculture.


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