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camalot
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Registered: 02/05/20
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Petri dish sectoring or not?
#26469908 - 02/05/20 08:58 AM (3 years, 11 months ago) |
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Hi everyone,
This is my first attempt to grow out mycelium on agar. I have used the "cloning technique".
Now, after 9 days of them "growing" on MEA agar in the dark between 24-27°C, I wonder what to do...
Shown in the photos are two dishes with lion's mane (LM1 en LM2) and two dishes with yellow oyster (YO1 en YO2). Both are cloned. The original mushroom were freshly bought from a local seller.
   
1/ So I wonder in general if it is needed to select a strain/isolate sectors when mycelium is grown out from a clone? As one should have 'a copy' of the original mushroom which is identical to its source? I read that people are doing this heavy isolating when growing form spores due to not having a monoculture. While cloning should result in a monoculture?
Questions related to the photos:
2/ For all dishes: is it needed to transfer any to another petri dish? Or can I directly inoculate with this?
3/ LM1 and LM2: for the lion's mane: LM2 started of as a 'blob' of almost silicon looking something (inner curly blue line on the photo). Since a few days it starts to show 'threads' (outer curly blue line on the photo). Is this normal? Does it mean that I should transfer it more until it grows directly in a more thready shape? Since today LM1 is also showing this behaviour.
4/ YO1 and YO2: for the yellow oysters: do they need a transfer? None of them look really uniform grown out. So am I correct that I can cut the most outer part and transfer? For example for YO1 the left clone, the sector between 12 and 1 o'clock? Soon they will start to "touch" each other ( as I did put 3 in a dish).
Thank you so much for all your help in advance! I am learning ;-)
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Forrester
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Registered: 02/05/13
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Re: Petri dish sectoring or not? [Re: camalot]
#26469958 - 02/05/20 09:38 AM (3 years, 11 months ago) |
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1. no, clone is good as is. No need to isolate anything other than isolating the culture from the contams.
2/3. I would transfer more on all of them. I don't know what that blobby crap is but it's not hericium, that thin wispy stuff coming out from that probably is. Get it out of there onto another dish until it's clean.
4. The oyster is probably fine but I'd take one transfer to make sure. This isn't from spore so it doesn't matter where you take the myc. from just as far from anything that looks like contamination as you can.
Nice job on the clear pics with labels, it makes it a lot easier to help you
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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katre
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Re: Petri dish sectoring or not? [Re: Forrester]
#27305302 - 05/12/21 07:05 AM (2 years, 8 months ago) |
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Hi, I would like to ask for help, too. I haven't found much information about sectoring lion's mane and I don't know how to go about it. I had a petri dish inoculated with liquid culture and it produced these little islands of mycelium (pic 1), which I read is normal. I then chose 2 areas and transferred them to new plates (pic 2 and 3). It grew nicely, but I am kinda stuck at this point...does it make sense to do further sectioning?
Thank you so much for any help.
 
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deadmandave
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Re: Petri dish sectoring or not? [Re: katre]
#27305618 - 05/12/21 11:14 AM (2 years, 8 months ago) |
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What's your goal? If it's just to grow some lions mane you don't need to do anything else except go to grains. If you're trying to generate a strain that's tolerant to higher temps or just looking for interesting characteristics you should start from a spore print and sector and grow out many different sectors.
Most likely the LC you have is not very genetically rich. I mean it's probably been cloned/plated many times before you got it but that's just a guess.
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Forrester
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Re: Petri dish sectoring or not? [Re: deadmandave]
#27312693 - 05/17/21 03:23 PM (2 years, 8 months ago) |
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Either way, you're not accomplishing anything by doing further transfers. The genetics have already been chosen so aside from making sure it's contamination free which you've already done, there's absolutely no reason to, you're just going to end up with the same thing.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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katre
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Re: Petri dish sectoring or not? [Re: Forrester]
#27355447 - 06/19/21 04:37 PM (2 years, 7 months ago) |
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Thank you both for your answers, that makes sense! So what about when the tissue culture is isolated from fruiting bodies and grown on agar? Is it possible/does it make sense to sector LM?
It doesn’t seem to be a common practice.
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deadmandave
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Re: Petri dish sectoring or not? [Re: katre]
#27355537 - 06/19/21 06:11 PM (2 years, 7 months ago) |
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Unless it's a multi spore grow there's really no reason to sector. The whole plate of a clone will have virtually the same genetic makeup, unless it came from MS.
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