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Offlinebeeker
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Agar, a badass clone, and bacteria
    #26611375 - 04/18/20 07:41 PM (3 years, 9 months ago)

Okay so after a year in the fridge, I pulled out a petri dish of a great clone and made a transfer to new agar, not really expecting to have much luck. To my surprise, the transfers to one of the plates is doing really well with the exception of some bacteria in one small segment near the mycelium. My question is this: If I start transferring rice grain size sections from the clean parts of the mycelium, I will essentially be creating isolates from my clone, correct? I'm trying to avoid that because to my understanding I would be separating all of the genetics of that clone out and would thus need to grow them all out to find the best isolate or just have a crapshoot and pick one or two hoping they have the majority of good traits that made the clone so awesome. Is there any way to conserve the genetics in their entirety of the clone itself. What's best to do at this stage? Am I missing something? Thanks for clarifying my understanding of this.


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Invisiblemushhead
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611385 - 04/18/20 07:46 PM (3 years, 9 months ago)

I'd attempt to save as much myc as possible and grow it back out.


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InvisibleTheStallionMang
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611442 - 04/18/20 08:15 PM (3 years, 9 months ago)

You can't isolate genetics from a clone because they're already from just one "parent".  Isolation is for when using spores.  In my understanding of it, you've got one set of genetics from your clone so you're just transferring agar to make sure you don't have contams.

Question for you now.. did your petri dishes start growing when you took them out or not until you transferred to another dish?  I recently threw out 6 dishes from the fridge because they didn't grow for 4 days after taking them out and I assumed they were toast.  They'd only been in the fridge about 6 months but maybe I needed to try a transfer :facepalm:


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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: TheStallionMang]
    #26611500 - 04/18/20 08:51 PM (3 years, 9 months ago)

Thanks mushhead, that's my plan. You'd suggest on big section to cut and transfer?

TSM, I'm pretty sure you can isolate from clones and spores. Clones sector out and are therefore not monocultures.

Petri dishes had bacteria from the year in the fridge and having to move them from house to house a few times.


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Invisiblemushhead
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611560 - 04/18/20 09:23 PM (3 years, 9 months ago)

truthfully it doesn't matter the size of the cut as long as you know you're getting good healthy myc. you can work around contams, and after a few transfers i'm sure you'll have a clean plate!

This is one of mine i'm currently working on here: https://www.shroomery.org/forums/showflat.php/Number/26604440 .


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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: mushhead]
    #26611619 - 04/18/20 09:51 PM (3 years, 9 months ago)

yeah, I'm sure I can get a clean plate on the next one. I've got a FH and good technique. I just find myself obsessing over getting the absolute best genetics and of course want that as soon as possible. Again and again this hobby is teaching me patience.


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InvisibleTheStallionMang
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611634 - 04/18/20 09:57 PM (3 years, 9 months ago)

Quote:

beeker said:
TSM, I'm pretty sure you can isolate from clones and spores. Clones sector out and are therefore not monocultures.




Ok follow my thought, if you start with one spore, grow it and eventually it grows shrooms, you've got the same genetic make up in each fruit (the spores in those fruits will have genetic variance but that isn't what we're talking about). A clone is called a clone because it has the same genes as the "parent" so if the petri dish was started with a cutting from a shroom, there is no genetic variance in that myc.


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InvisibleTheStallionMang
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611644 - 04/18/20 10:04 PM (3 years, 9 months ago)

Quote:

beeker said:
Petri dishes had bacteria from the year in the fridge and having to move them from house to house a few times.




did your petri dishes start growing when you took them out or not until you transferred to another dish?

I'm trying to know if I wasted some dishes or not.  Did you keep the dish from the fridge at room temp long enough to know if it was growing before you took a cutting?


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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611654 - 04/18/20 10:09 PM (3 years, 9 months ago)

here's a pic of what i'm working with from both sides. I find the backlight underside to be most helpful in identifying bacteria



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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611662 - 04/18/20 10:11 PM (3 years, 9 months ago)

pic 2



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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: TheStallionMang]
    #26611671 - 04/18/20 10:15 PM (3 years, 9 months ago)

I transferred after about a day. They hadn't started growing out yet, mainly because they had no space left on the plate.


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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: TheStallionMang]
    #26611679 - 04/18/20 10:17 PM (3 years, 9 months ago)

I was under the impression you couldn't start with one spore. I believe you start with two spores before any mycelium will appear, true?


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InvisibleTheStallionMang
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611695 - 04/18/20 10:26 PM (3 years, 9 months ago)

Ok thanks for answering that.

TBH I don't know if you can start from one spore or not.  I was using it as example. I seems like it should work but I'm not the most knowledgeable about it. It'd be pretty darn hard to manipulate just one spore into agar anyway.

The growth in you're new dishes look pretty good


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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: TheStallionMang]
    #26611743 - 04/18/20 11:03 PM (3 years, 9 months ago)

Quote:

TheStallionMang said:
Ok thanks for answering that.

TBH I don't know if you can start from one spore or not. 

The growth in you're new dishes look pretty good





Thanks, yeah you have to get two spores to mate before you can get any growth.


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InvisibleTheStallionMang
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Re: Agar, a badass clone, and bacteria [Re: beeker]
    #26611750 - 04/18/20 11:11 PM (3 years, 9 months ago)

So I see you're pretty new here, have no ratings and haven't left any. So in case you didn't know, we can rate each other and then others can check ratings when considering if they should take user's advice and such.
I just gave you your first rating :thumbup:


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OfflineMachiavelliavore
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Re: Agar, a badass clone, and bacteria [Re: TheStallionMang]
    #26611824 - 04/19/20 12:15 AM (3 years, 9 months ago)

Quote:

TheStallionMang said:
Ok follow my thought, if you start with one spore, grow it and eventually it grows shrooms, you've got the same genetic make up in each fruit (the spores in those fruits will have genetic variance but that isn't what we're talking about). A clone is called a clone because it has the same genes as the "parent" so if the petri dish was started with a cutting from a shroom, there is no genetic variance in that myc.




Many pairs of spores form the many strains in a multispore grow, and many strains form the fruits.  If there was no diversity in the strains that formed the fruit, you wouldn't get sectoring from a clone.  Supposedly the exchange of nuclei between cells makes it such that most of those strains will be very similar, especially as time passes.  If your clone has already seen a few transfers in its lifetime, much of the diversity that once existed is probably already gone from both growing and exchanging genetics and from accidental isolation, and not worth worrying about.

I'm not sure to what degree different isolates of a single clone have been tested against eachother by members here.  Commercial strains usually come from MS isolates afaik, so probably not much data there.

I think for some reason the actual term isolate may not be preferred in the context of clones, but I've never been clear on why.


--------------------


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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InvisibleTheStallionMang
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Re: Agar, a badass clone, and bacteria [Re: Machiavelliavore]
    #26611924 - 04/19/20 01:56 AM (3 years, 9 months ago)

Interesting, I'd be curious to know if anyone has tested different isolates from a clone to compare results. Surely someone here has...

I recognize you by your signature, which is perfect for these "shelter in place" days


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Offlinebeeker
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Re: Agar, a badass clone, and bacteria [Re: TheStallionMang]
    #26612422 - 04/19/20 08:44 AM (3 years, 9 months ago)

Quote:

TheStallionMang said:
Interesting, I'd be curious to know if anyone has tested different isolates from a clone to compare results. Surely someone here has...





That's a better way of asking for exactly the info I'm looking for. :smile:


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Invisiblemushhead
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Re: Agar, a badass clone, and bacteria [Re: beeker] * 1
    #26612535 - 04/19/20 09:28 AM (3 years, 9 months ago)

Quote:

beeker said:
Quote:

TheStallionMang said:
Interesting, I'd be curious to know if anyone has tested different isolates from a clone to compare results. Surely someone here has...





That's a better way of asking for exactly the info I'm looking for. :smile:



each spore is called a monokaryote, when they "fuse" they're called dikaryotes, you can isolate singular monokaryotes to germinate specific genes. Take a clone or an isolated myc strand from plate, grow it to fruition and then breed monokaryotes into dikaryotes to attempt to isolate the genetics you want. Hope this helped.


--------------------
Meditation Principles

Silence: Giving you room to listen / Stillness: Giving you room to feel / Spaciousness: Just giving you room


Edited by mushhead (04/19/20 11:46 AM)


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