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Offlinefrikiriki
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Genetics, Isolating, Cloning, A Few Questions...
    #27288248 - 05/01/21 04:34 AM (1 year, 8 months 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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InvisiblefahtsterM
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: frikiriki]
    #27297999 - 05/07/21 11:00 AM (1 year, 8 months 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.

Faht


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Offlinefrikiriki
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: fahtster]
    #27303572 - 05/11/21 06:26 AM (1 year, 8 months ago)

Thank you! I will continue cloning clones... and I will now apply this new generation information to my practice. Also, regarding the theory that there are multiple genetics within a clone, yes this does seem very true. That is why I grab several fruit bodies from a single grow, and take multiple tissue samples from each. I have another question as well, if you don't mind. Do spores get better/more refined through each generation if you know what you are doing with the genetics?


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InvisiblefahtsterM
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: frikiriki]
    #27303590 - 05/11/21 06:49 AM (1 year, 8 months ago)

Well if you find a phenotype you like from an ms grow, you can keep selecting that pheno from each new generation from spore and eventually you narrow the genetics down so that pheno will be the only one expressed from multispore.. for the most part.  Environmental factors can play a role to what a fruit looks like but the majority of fruits will look like that one you kept choosing.

Faht


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Offlinefrikiriki
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: fahtster]
    #27305472 - 05/12/21 06:29 PM (1 year, 8 months ago)

Wow! Awesome. I appreciate the information, thanks again!


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OfflineFrugalFungi
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: fahtster]
    #27312802 - 05/18/21 02:02 AM (1 year, 8 months ago)

Quote:

fahtster said:
Well if you find a phenotype you like from an ms grow, you can keep selecting that pheno from each new generation from spore and eventually you narrow the genetics down so that pheno will be the only one expressed from multispore.. for the most part.  Environmental factors can play a role to what a fruit looks like but the majority of fruits will look like that one you kept choosing.

Faht




Just what I was looking for thanks :smile:


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Offlineelunicotomas
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: fahtster]
    #27313053 - 05/18/21 06:55 AM (1 year, 8 months ago)

Quote:

fahtster said:
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.





I've read somewhere about this and I can't find it now, the thing is that if you keep cloning your fruits eventually you won't get fruits right?


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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: elunicotomas]
    #27313061 - 05/18/21 07:08 AM (1 year, 8 months ago)

It takes a good amount of “reclones”.. I’d imagine you can do it at least a few times before you start noticing anything depending on how old the culture is

Faht


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Offlineelunicotomas
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Re: Genetics, Isolating, Cloning, A Few Questions... [Re: fahtster]
    #27313307 - 05/18/21 03:07 PM (1 year, 8 months ago)

Quote:

fahtster said:
It takes a good amount of “reclones”.. I’d imagine you can do it at least a few times before you start noticing anything depending on how old the culture is

Faht




I cloned a big mushroom i got last month, made 1 or 2 transfers, and inoculated some grain jars which I'll be using in about a week or so, so the culture is not old.... I'll see what comes from this and future cloning, thanks!


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