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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Questions on cloning in agar
    #25514486 -

I'm taking the agar plunge shortly and will be seeking to clone my best and brightest.  However, since this would be initially from an MS syringe I'd think it's guaranteed to grow out into a lot of separate sectors. 

My thought process is that if I transfer a single sector it is only part of the genetics I was seeking to recreate.  So, for a true clone would I need to take a portion of each sector?  Perhaps concentric rings to multiple plates?  It may be as simple as waiting for it fruit and transferring the pin but I hit the same mental wall - if I'm taking wedges for grain they'd only represent the myc from that wedge.

Apologies if I'm missing something obvious here - just dipping my toes in the agar pool.


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The Lazy Man's Teks

Agar as easy as 1-2-3!
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Re: Questions on cloning in agar [Re: trip_hazard] * 1
    #25514518 -

When you clone, you're growing out the genetic variation that particular fruit holds, as much as it holds. Each series of sector isolation from said clone is refining your choice. Unfortunately you won't know what fruit phenotypes you'll have unless you grow it out. I prefer to clone, isolate a few times ( random selection ) then grow out again. Repeat until your growth is consistent or satisfactory. Have patience and persistence because it takes a while from ms. Make backup cultures throughout your transfers, keep organized and document the ancestry. And have great diligence when judging the cleanliness of your cultures


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Re: Questions on cloning in agar [Re: Noisecontrol]
    #25514539 -

Ah - I think it is becoming more clear.  I've got some jars from MS that should be ready in the next week or so.  My plan is to take my star pupil(s) from that grow for cloning.  Based on what you've stated, and some other reading, the genetics will be the same but each sector is an expression of phenotype and it's 'luck of the draw' on what each phenotype grows into when birthed.

Based on this, is it fair to say there is no such thing as a perfect phenotypical clone?


--------------------
:mushroom2::mushroom2::mushroom2: :mushroom2::mushroom2::mushroom2:


The Lazy Man's Teks

Agar as easy as 1-2-3!
Trip's Sous Vide Rye Prep
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Making shoeboxes


Crucial Reference Threads

Proper Surface Moisture / When to Increase FAE

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Re: Questions on cloning in agar [Re: trip_hazard]
    #25515066 -

No and no, the genetics will not be the same because you're not at a static monoculture yet. And this is why it is referred to as isolation. When your culture expands radially on the plate, each genetic possibility is there, clustered within the space available. Each time you "isolate" a "sector" of the culture, the genetics contained therein are given space to spread out on a new plate. This process is repeated until there is little to no genetic drift. Think of your starting culture, either from spore or clone, as a really high number expressed from 1 to that number clockwise like a clock (representing the possibilities of growth), each time you take a chunk of that clock, those numbers selected have more space to spread out on their own clock. It's process of elimination.


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Re: Questions on cloning in agar [Re: Noisecontrol]
    #25515431 -

Shit - more confused now after reading this thread.


--------------------
:mushroom2::mushroom2::mushroom2: :mushroom2::mushroom2::mushroom2:


The Lazy Man's Teks

Agar as easy as 1-2-3!
Trip's Sous Vide Rye Prep
Bod's easy AF bucket tek
Making shoeboxes


Crucial Reference Threads

Proper Surface Moisture / When to Increase FAE

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Re: Questions on cloning in agar [Re: trip_hazard] * 1
    #25515594 -

If you have fruits from MS coming, get a clone of your biggest fastest fruit.

I recommend biopsy of a small piece of tissue from the lower/middle of the stem. You can use a scalpel or a needle to get a tiny tiny piece. Needle poke helps with contams. Expect a transfer or two to remove bacteria.

Thats a clone. You can make multiple plates or a slant and store it for re-use if you like it.

Using spores from any fruit will give you unlimited genetic variation even between varieties like GT b+, etc.

Isolation is to remove bacteria #1 and make a monoculture for efficient colonization #2 and #2 isn't required. Any isolation from MS will give you a random isolation you know nothing about. If you grow it, it may be complete shit. You don't know. It could also be awesome and prolific.

I'd recommend #1 grow from MS, then #2 clone a favorite. That way you isolate something you already know. Remove bacteria and push on. Use those new spores for MS and play with isolates in your free time if you'd like.

A great method is letting your MS agar plate grow out 100% and clone the tiny fruit it produces. That will be your fastest colonizer(on agar, so use your grain water), and a proven fruiter.

LMK if you need any more info. Someone correct me if im wrong.


I take no credit for any of this info. Mostly BOD probably but its a collection of many teks from many people. I just research this site and think about it before trying something stupid.

Edited by mushfuss (10/05/18 06:52 PM)

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Re: Questions on cloning in agar [Re: Noisecontrol]
    #25515628 -

Noisecontrol said:
No and no, the genetics will not be the same because you're not at a static monoculture yet. And this is why it is referred to as isolation. When your culture expands radially on the plate, each genetic possibility is there, clustered within the space available. Each time you "isolate" a "sector" of the culture, the genetics contained therein are given space to spread out on a new plate. This process is repeated until there is little to no genetic drift. Think of your starting culture, either from spore or clone, as a really high number expressed from 1 to that number clockwise like a clock (representing the possibilities of growth), each time you take a chunk of that clock, those numbers selected have more space to spread out on their own clock. It's process of elimination.



Does a biopsy clone from a fruit still have lots of genetic variation?

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Re: Questions on cloning in agar [Re: mushfuss]
    #25515725 -

A clone from MS still contains multiple strains typically, yes...


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Re: Questions on cloning in agar [Re: Caps McGee]
    #25515739 -

Caps McGee said:
A clone from MS still contains multiple strains typically, yes...


Interesting. I had assumed that they fruit independently producing the variation. I'm still at the first brf cake stage, no hands on experience.

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Re: Questions on cloning in agar [Re: Caps McGee]
    #25515740 -

Also, I suggest running several clones to find a favorite, and slant that one, then further isolate the culture through cloning and transfer... keep the original clone culture slanted to go back to, in case isolation ends up bunk... IME, the clone culture  (which is a monoculture,  even with multiple strains) performs better than most isolates, narrowing genetics is a crap shoot and can take years to find 'the one'... your current focus is cleanliness, obtaining and transferring a clean culture...


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Re: Questions on cloning in agar [Re: Caps McGee]
    #25515745 -

So clones produce monocultures, not isolates. Isolates are single strain, and a monoculture is multi-strain but with identical growth?

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Re: Questions on cloning in agar [Re: mushfuss]
    #25515757 -

Monoculture, (from my understanding )is a loose term with a few definitions... one sense is as the name implies, one culture... meaning only mucelium, a clean culture... but generally speaking in our minds it refers to a culture acting as one,  as it came from only one fruit,  it's only one culture, be that it may be several strains working together as one


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Re: Questions on cloning in agar [Re: Caps McGee]
    #25515763 -

Awesome, thanks. mono culture is a much better goal than isolate unless you have something special to start with is my take-away.

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Re: Questions on cloning in agar [Re: mushfuss]
    #25515764 -

Caps McGee said:
A clone from MS still contains multiple strains typically, yes...


mushfuss said:
Isolation is to remove bacteria #1 and make a monoculture for efficient colonization #2 and #2 isn't required. Any isolation from MS will give you a random isolation you know nothing about. If you grow it, it may be complete shit. You don't know. It could also be awesome and prolific.




Precisely - not trying to isolate.  That's what led to my question - if I'm picking my favorite and cloning it I want it as close to the original as possible.  Would it be possible to capture all the strains of the source fruit?


--------------------
:mushroom2::mushroom2::mushroom2: :mushroom2::mushroom2::mushroom2:


The Lazy Man's Teks

Agar as easy as 1-2-3!
Trip's Sous Vide Rye Prep
Bod's easy AF bucket tek
Making shoeboxes


Crucial Reference Threads

Proper Surface Moisture / When to Increase FAE

Extras: Filter Print Post Top
Re: Questions on cloning in agar [Re: trip_hazard]
    #25515769 -

That should produce the full monoculture of that specific fruit if they all recover. You can't get the genetics from the parent of your 3rd party spore source.

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Re: Questions on cloning in agar [Re: mushfuss]
    #25515775 -

No... you could take several samples from the same fruit and all grow out different


--------------------
🍄💚😎

Ask Caps! (My Q&A Thread)
Caps' Comprehensive Cultivation Tek List

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Re: Questions on cloning in agar [Re: mushfuss]
    #25515781 -

mushfuss said:
That should produce the full monoculture of that specific fruit if they all recover.


That's what I'm after - thus the question about using concentric rings of an agar plate instead of wedges.


--------------------
:mushroom2::mushroom2::mushroom2: :mushroom2::mushroom2::mushroom2:


The Lazy Man's Teks

Agar as easy as 1-2-3!
Trip's Sous Vide Rye Prep
Bod's easy AF bucket tek
Making shoeboxes


Crucial Reference Threads

Proper Surface Moisture / When to Increase FAE

Extras: Filter Print Post Top
Re: Questions on cloning in agar [Re: Caps McGee]
    #25515791 -

Caps McGee said:
No... you could take several samples from the same fruit and all grow out different


Bugger.  So no way to duplicate a set of phenotypes?


--------------------
:mushroom2::mushroom2::mushroom2: :mushroom2::mushroom2::mushroom2:


The Lazy Man's Teks

Agar as easy as 1-2-3!
Trip's Sous Vide Rye Prep
Bod's easy AF bucket tek
Making shoeboxes


Crucial Reference Threads

Proper Surface Moisture / When to Increase FAE

Extras: Filter Print Post Top
Re: Questions on cloning in agar [Re: trip_hazard]
    #25515793 -

trip_hazard said:
mushfuss said:
That should produce the full monoculture of that specific fruit if they all recover.


That's what I'm after - thus the question about using concentric rings of an agar plate instead of wedges.


Transfer the outermost circle to agar. Wedges are for grain inoculation. There are parts of the outer circle that are mostly isolated and mostly variated. Grab some good looking stuff with a tiny tiny half grain sized bit to lower variation. Agar biopsy with a needle would do great as well.


Thanks for clarifying, caps.


Maybe I should hold off on the advice giving :smile:

Edited by mushfuss (10/05/18 07:55 PM)

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Re: Questions on cloning in agar [Re: mushfuss]
    #25515805 -

You can get lucky, and some traits may be more consistent than others... thing about growing a bunch of mushrooms looking for the perfect isolate, is that you've got A BUNCH OF FUCKING MUSHROOMS!


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Ask Caps! (My Q&A Thread)
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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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