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InvisiblemushboyMDiscord
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Re: Cloning from a clone culture [Re: Mad Season]
    #23937574 - 12/18/16 10:02 AM (7 years, 1 month ago)

i have one going now. its from an invitro pin off the original cloned tissue.

growth has been good...

am i assuming there are equal chances of 'isolating' bad genetics vs good since
its only a pin and i dont know its full characteristics?


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InvisibleMad Season
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Re: Cloning from a clone culture [Re: mushboy]
    #23937578 - 12/18/16 10:03 AM (7 years, 1 month ago)

I wouldnt say equal chances, but there is a chance you can see both good and bad characteristics.


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Invisibler.lutece
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Re: Cloning from a clone culture [Re: Mad Season]
    #23937579 - 12/18/16 10:03 AM (7 years, 1 month ago)

I can't help but love questions about strains and isolating. Every answer brings up two questions for me.

When we transfer, we take a very small sample of mycelium. What happens if we take a large sample? Does it increase the odds of contamination?

Also (and this may be getting into realms that we need more advanced equipment to adequately study), if we take samples from two nonadjacent sectors in a cloned subvariety, will those sectors be compatible when reintroduced in a new medium? Or would it be akin to introducing two strains that were isolated individually?


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Offlineenlightenment
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Re: Cloning from a clone culture [Re: r.lutece]
    #23937592 - 12/18/16 10:07 AM (7 years, 1 month ago)

Quote:

When we transfer, we take a very small sample of mycelium. What happens if we take a large sample? Does it increase the odds of contamination?




It increases the chance that you transfer another unwanted sector.

I pass the second question to Mad. :laugh:


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InvisibleMad Season
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Re: Cloning from a clone culture [Re: enlightenment]
    #23937611 - 12/18/16 10:13 AM (7 years, 1 month ago)

Theres a possibility it can merge and have anastomosis, but there's also a possibility they won't. Regardless tho they're both cubensis and are compatible with each other, they will still easily transfer water and nutrients between them, regardless of anastomosis


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InvisibleMush 4 Brains
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Re: Cloning from a clone culture [Re: PinPornProducer]
    #23937620 - 12/18/16 10:16 AM (7 years, 1 month ago)

Quote:

PinPornProducer said:
Quote:

cronicr said:
You can clone from a clone it just sounds silly...you should already have it on agar.



A clone still has multiple genetics though, so wouldn't cloning a clone reduce those genetics even more? Kinda like isolating?



I sort of doubt it. When you clone a clone, all youre doing is expanding mycelium on to another medium.

It probably would experience genetic drift and preform poorly as a result because of how many generations old it is. Just the fungus taking another form, from fruit to mycelium and back to fruit again.

I cant picture you isolating or narrowing down any of the sub-strains by cloning a clone.


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Invisibler.lutece
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Re: Cloning from a clone culture [Re: Mad Season]
    #23937624 - 12/18/16 10:19 AM (7 years, 1 month ago)

Quote:

Mad Season said:
Theres a possibility it can merge and have anastomosis, but there's also a possibility they won't. Regardless tho they're both cubensis and are compatible with each other, they will still easily transfer water and nutrients between them, regardless of anastomosis




Makes sense. I wasn't sure if there would be increased compatibility between strains that originated from the same clone.


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But one does not undertake an experiment knowing one HAS failed.


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OfflinecronicrFacebook
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Re: Cloning from a clone culture [Re: Mush 4 Brains]
    #23938084 - 12/18/16 01:37 PM (7 years, 1 month ago)

Quote:

Mush 4 Brains said:
Quote:

PinPornProducer said:
Quote:

cronicr said:
You can clone from a clone it just sounds silly...you should already have it on agar.



A clone still has multiple genetics though, so wouldn't cloning a clone reduce those genetics even more? Kinda like isolating?



I sort of doubt it. When you clone a clone, all youre doing is expanding mycelium on to another medium.

It probably would experience genetic drift and preform poorly as a result because of how many generations old it is. Just the fungus taking another form, from fruit to mycelium and back to fruit again.

I cant picture you isolating or narrowing down any of the sub-strains by cloning a clone.



This is semi true but gen are defined by spores not expansion..so say you grew a cake..took a clone and grew that on a qt of rye...you still have very little cell division and likely still have more then one strain so taking another clone wouldn't really hurt anything but again the smart one still has the original plate which would be easier n smarter to work with...i am yet to see a real diff with cubes weather you isolate from a clone or try keep it whole.

The problems would come if you were the guy who took that clone..made a master and expanded it out a bunch of times then grew it and tried to clone again.


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InvisibleMad Season
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Re: Cloning from a clone culture [Re: cronicr]
    #23938327 - 12/18/16 02:56 PM (7 years, 1 month ago)

Correct me if I'm wrong but isolating strains regardless of agar transfers or clones of clones are all forms genetic drifting.

And apparently in this thread PF had non-fruiters in a sample beginning with a clone. Seems vastly different from a clone, and I've had many times where clones with a couple transfers performed worse than their MS counterparts, and vice versa obviously.

Shit's all a crapshoot lol. I do agree tho that continuously expanding and cloning would hinder performance a lot more.


--------------------
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OfflinecronicrFacebook
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Re: Cloning from a clone culture [Re: Mad Season]
    #23938344 - 12/18/16 03:02 PM (7 years, 1 month ago)

Yep all clones.and isolates need testing for good reason..i myself have run into wierd things where i clone a cluster and it pops out a bunch of single fruits...ya just never fucking know lol.


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InvisibleMush 4 Brains
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Re: Cloning from a clone culture [Re: cronicr]
    #23938368 - 12/18/16 03:11 PM (7 years, 1 month ago)

Quote:

cronicr said:
Quote:

Mush 4 Brains said:
Quote:

PinPornProducer said:
Quote:

cronicr said:
You can clone from a clone it just sounds silly...you should already have it on agar.



A clone still has multiple genetics though, so wouldn't cloning a clone reduce those genetics even more? Kinda like isolating?



I sort of doubt it. When you clone a clone, all youre doing is expanding mycelium on to another medium.

It probably would experience genetic drift and preform poorly as a result because of how many generations old it is. Just the fungus taking another form, from fruit to mycelium and back to fruit again.

I cant picture you isolating or narrowing down any of the sub-strains by cloning a clone.



This is semi true but gen are defined by spores not expansion..so say you grew a cake..took a clone and grew that on a qt of rye...you still have very little cell division and likely still have more then one strain so taking another clone wouldn't really hurt anything but again the smart one still has the original plate which would be easier n smarter to work with...i am yet to see a real diff with cubes weather you isolate from a clone or try keep it whole.

The problems would come if you were the guy who took that clone..made a master and expanded it out a bunch of times then grew it and tried to clone again.



Generations, since ive joined this website have always referred to mycelium transfers. Like each g2g is a generation. Look it up,  you'll find RR referring to generations as mycelium transfers like g2gs etc.

As i recall the consensus was you dont want to go past 5 or so gens. I specifically recall RR saying you can get away with going past 5 or so generations provided the mycelium is young, robust and you move quickly. But nomenclature in cultivation is kind of all over the place. Different people use different words to describe this or that.

Also thats pretty much what i was referring to, using the clone of a clone to start a new grow/master.


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InvisiblePastywhyteMDiscord
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Re: Cloning from a clone culture [Re: Mush 4 Brains]
    #23938384 - 12/18/16 03:16 PM (7 years, 1 month ago)

A G2G is a P value as is a 90mm plate grown out. That is a reference to expansion. I have never seen an expansion referred to as a generation. A generation as I understand it is from spore to fruit to spore.


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InvisibleMush 4 Brains
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Re: Cloning from a clone culture [Re: Pastywhyte]
    #23938514 - 12/18/16 03:55 PM (7 years, 1 month ago)

I know info changes, words/descriptions change, ideology and of course techniques through the years. But im 100% certain that I'm correct. I'll do some searching and dig up an example by RR and or others.

Transfers on agar are excluded from this. It was specifically and almost exclusively in reference to g2g. For example ms to grain jar is gen 1, then g2g from  that grain is gen 2 and so on for each transfer.

Im sure some old hands here can back me up on this. I dont stop into cultivation forum much anymore but everytime i do there seems to be new methodology/ideology and nomenclature


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InvisibleMush 4 Brains
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Re: Cloning from a clone culture [Re: Mush 4 Brains]
    #23938528 - 12/18/16 04:02 PM (7 years, 1 month ago)

Ok so i found this which backs up what you guys are saying. Even RR said its incorrect nomenclature to call transfers generations however ive been around long enough to see RR stick his foot in his mouth. That is to say he used to say x but later says y. All i know is people at one time did refer to each g2g as a generation.

https://www.shroomery.org/forums/showflat.php/Number/15867731#15867731


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Re: Cloning from a clone culture [Re: Mush 4 Brains]
    #23938531 - 12/18/16 04:03 PM (7 years, 1 month ago)

I think it was stamets who used to call them gens?


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InvisiblePastywhyteMDiscord
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Re: Cloning from a clone culture [Re: Mush 4 Brains] * 1
    #23938538 - 12/18/16 04:04 PM (7 years, 1 month ago)

Here is a great one from workman who's work I have much respect for. Note his use of generation.


Quote:

Workman said:
Mushrooom genetics are a little strange since a single mushroom produces spores that can then act as both parents for a new mycelium.  Essentially, you are selfing or inbreeding each time you do a multispore grow.

Now consider a wild collection of Psilocybe cubensis with a high heterozygosity.  This basically means that most or all of each pair of genes in the mushroom are different from each other.  Its the same gene location with the same basic function, but different versions.  For example, if there is a single gene for height, you might have a version that gives short mushrooms and a version that gives tall mushrooms.  If heterozygosity is high, you have one of each which may result in medium mushrooms unless one of the height genes is dominant.

Now, when you do multispore from a single mushroom you randomly get a mix of all the genes.  Sticking to our height gene example, you could get two short copies, two tall copies or one of each.  Obviously the strains with two short copies will be short and the ones with two tall copies will be tall. 

Lets say we liked the short mushrooms so we saved that one and took a spore print for later.  In this example the tall version of the height gene is lost to later generations.  There is a net loss of heterozygosity.  Over the entire genome the loss is about 50% per generation.

So mathematically we can figure out how many sequential multispore generations we need until the heterozygosity is reduced to an insignificant level and the strain is stable even from multispore.

Starting with a presumably high (~100%) heterozygosity from a wild collection.  In reality, the heterozygosity is probably lower than 100%, but its an easy number to start with.

100% wild print
50% 1st generation from wild print
25% 2nd generation from 1st generation print
12.5% 3rd generation.....
6.25% 4th generation.....
3.12% 5th generation.....
1.56% 6th generation.....
0.78% 7th generation.....

You can see that the heterozygosity drops off quickly in the first few generations and is less than 1% after the 6th generation.  This highlights the importance of choosing the best traits early on when there are more to choose from.  Attempting to isolate traits in well established strains results in only minimal improvements unless spontaneous mutations increase the heterozygosity in a positive way (rare).

In summary:

Popular classic strains in circulation have all been grown well beyond 6 generations and are relatively stable from multispore with little need for isolation.

New strains, from wild material or cross breeding between different strains of the same species, can be stabilized fairly quickly with 6 or 7 generations of sequential multispore grows.

Selection is most important early in the process and if good genes are bred out, they are gone forever.  Archiving original or early generation prints is recommended for preserving heterozygosity for later selective breeding.  Continuous isolation of a bad strain with hopes of significant improvement is futile.

Does that help?





https://www.shroomery.org/forums/showflat.php/Number/10242476#10242476


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Invisibleblindingleaf
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Re: Cloning from a clone culture [Re: Pastywhyte]
    #23938555 - 12/18/16 04:11 PM (7 years, 1 month ago)

I think it's 2 kind of uses of the word "generation"-one has to do with genetics and stabikization and the other has to do with grain expansion.  it's normal to say "I went to G3 before going to bulk"


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Invisibleblindingleaf
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Re: Cloning from a clone culture [Re: blindingleaf]
    #23938559 - 12/18/16 04:12 PM (7 years, 1 month ago)

Oh wait...I just realized the g could stand for grain too :facepalm:


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InvisibleMush 4 Brains
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Re: Cloning from a clone culture [Re: blindingleaf]
    #23938677 - 12/18/16 05:00 PM (7 years, 1 month ago)

Interesting stuff Pasty. I have a lot of respect for the old school guys.. Ohmatic, MonsterMitch, agar, Largedose, Hippiechick to name a few. Rr i always found to be pretentious and i never cared for how he would insert his shitty political opinions into threads but no doubt the man knows his shit. Whatever happened to him anyway? He's still listed as a mod here, no?

But yeah all those guys really wrote the book on cultivation essentially starting from scratch. Plenty of them had a science background. Pretty sure RR said he used to work for Microsoft

You know it really doesnt make much sense to refer to g2gs as generations. A bit misleading


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Re: Cloning from a clone culture [Re: Mush 4 Brains]
    #23938684 - 12/18/16 05:03 PM (7 years, 1 month ago)

Rr stopped growing mushrooms and started growing weed up until his wife passed away..he only did both for her.


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