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Offlinefrikiriki
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Genetics, Isolating, Cloning, A Few Questions...
    #27288248 - 04/30/21 09:34 PM (6 days, 12 hours ago)

Hey all! I've been searching through some posts but I still have questions regarding what is FACTUAL with genetics. I have 2 close friends I am growing with, and am the laboratory personnel if you will. I extensively take notes, study, and label my petri dishes. I have recently encountered some divided opinions.

1. "Do not clone a clone."

2. "If you clone a clone, that becomes the 2nd generation."
   
3. ^^^ Opposing thought, no, a 2nd generation is when you grow out a clone, and start spores from this clone.

4. "If you grow out a tissue sample in a petri dish, and isolate from said dish onto a new one, the resulting dish will be genetically the same."

5. ^^^ Opposing thought, insisting that no, they will not be the same, you have separated/altered genetics.


Can anyone help to clear up exactly how you would regard generations, genetics, and so on and so forth?

Slightly unrelated, but I have done/found some things that differ from the regular methods, and I would like to share. 

I prefer to use sterile cotton swabs instead of liquid culture. I shred off the tip of the swab onto the agar. The spores begin growing much faster this way. The cotton provides a fantastic growing medium in this early stage, whereas the liquid is too watery and doesn't settle quite right.

I let spore dishes grow out a little longer than usual, allowing hyphae knots to form. If you isolate 1 hyphae knot, it grows out beautifully. 1 isolation in comparison to the multiples I was doing to get good rhizomorph mycelium.

If the spore dish is free of contaminates and stress indications, the whole dish goes to grain as a multi-spore grow.

Do lots and lots of individual spore dishes. There will be a lot of variety.

I have been cloning clones several times over and have found that you see more consistent, fuller, flushes. Yes, senescence does begin to occur after about 3-4 repetitions. (I gather spores throughout these grows to begin fresh.)

These repetitions are what I previously thought where considered "generations". Now I don't know what to think.

I have isolated a sector from a tissue sample from a clone of a clone, and it resulted in a completely different looking bin. Now I lean more towards the thought that isolating from a tissue sample DOES INDEED alter genetic makeup.

Help me, help you, help everybody in the name of mycology genetics research!

Thank you, sorry for the length, but I got some shit to say!


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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: frikiriki]
    #27297999 - 05/07/21 04:00 AM (6 hours, 15 minutes ago)

1. You can clone a clone.. I’ve done it a few times to save the culture with no adverse effects.  If you keep doing it over and over, eventually you’ll run into problems.
2. & 3. Yes, the next generation comes from starting from spores.. clone of a clone is just that.. a clone.

4. & 5. If you pull tissue from a ms plate, you’re isolating genetics from a mix of multiple genetics... you usually have to do many transfers like that to isolate down to a single genetic line or monoculture.  If you think that it only takes two spores mating to produce a monoculture and there’s thousands of spores in a very small amount of water, you can see why it takes so much to isolate down to that when all those spores germinate and find mating pairs.

There’s debate that there’s many genetic lines in even one cloned fruit body.. could be why you’re seeing different growth from a clone of a clone and not because there’s altering of genetic material, but mutations aren’t uncommon in fungi.

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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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