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Offlinekoma23
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Need some agar sectoring suggestions * 2
    #28647374 - 02/05/24 09:10 AM (3 months, 10 days ago)

Hi! First time using agar and decided to make some clones. I let them grew for some time, as I was interested in how the mycelium would expand, specifically, was interested in these long strands, as I haven't seen much agar that looked liked this. Which brings me to the questions;

- please advise on how to go about selecting sectors to transfer. Should I isolate these long strands or just ignore and select a wider sector?

- as I particularly liked the fruits I selected for cloning, wouldn't it be better to not micro select only specific sectors - as those wont be clones anymore but potentially totally different fruits - and rather use all the mycelium? I'm a bit confused on this part, as I used to clone by throwing tissue samples directly in the LC (yes I know ...) which did produce rather the same looking fruit ..

Appreciate any pointers!





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Invisiblejoze
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Re: Need some agar sectoring suggestions [Re: koma23] * 2
    #28647654 - 02/05/24 01:37 PM (3 months, 9 days ago)

Plate #3 looks a bit blue -- is that from bruising, or do you have some mold growing in there too?

You've got some nice rhizomorphic growth, definitely the most "aggressive" growth is on plates #1 and #2. However, just because something looks good on a plate, doesn't mean it's going to produce great fruits. When you're selecting for isolates and then fruiting them, it's always a gamble what you'll get. So I just take a bunch of different transfers and fruit them all, and see what I get.

I would take 3 or 4 transfers each from plates #1 and #2, that all looks pretty good. Plate #3 is a little bit suspect -- if it's just bruising, and not mold, then I'd take a couple transfers from 12 o'clock on that plate. On plate #4, I would take transfers from 1 o'clock and 4 o'clock.

As for the transfers themselves, I take a piece barely larger than a grain of rice. I essentially cut out the smallest piece I can manage. I do three cuts to cut a tiny little triangle, then stab it with the tip of my scalpel and scoop it out.


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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: joze] * 1
    #28647751 - 02/05/24 03:10 PM (3 months, 9 days ago)

Quote:

joze said:
Plate #3 looks a bit blue -- is that from bruising, or do you have some mold growing in there too?





Hey, really appreciate your input!

The "blue" on plate 3 is just the loss of color of the agar (I presume due to mycelium eating it up), so the black table can be seen through more clearly. Ie. it's not blue - just more transparent.

Quote:

joze said:
When you're selecting for isolates and then fruiting them, it's always a gamble what you'll get.




Yeah, I figure as much. But what about if you just want to clone and not isolate, what do you do then with agar?

Quote:

joze said:
I would take 3 or 4 transfers each from plates #1 and #2, that all looks pretty good. Plate #3 is a little bit suspect -- if it's just bruising, and not mold, then I'd take a couple transfers from 12 o'clock on that plate. On plate #4, I would take transfers from 1 o'clock and 4 o'clock.

As for the transfers themselves, I take a piece barely larger than a grain of rice. I essentially cut out the smallest piece I can manage. I do three cuts to cut a tiny little triangle, then stab it with the tip of my scalpel and scoop it out.




Ok, I see you're targeting what looks to be the most fast/strong growth.

Is this how you'd take out the slices?


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Invisibleelasticaltiger
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Re: Need some agar sectoring suggestions [Re: koma23] * 1
    #28647763 - 02/05/24 03:16 PM (3 months, 9 days ago)

Very beautiful ropes in those first two pictures.

Personally I would just take a few transfers from the rope ends of either of those first two dishes. Ignore 3rd dish.

If you're hell bent on transfering from the last dish that triangle you drew is how I'd do it.


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OfflineYahra
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Re: Need some agar sectoring suggestions [Re: elasticaltiger]
    #28647781 - 02/05/24 03:36 PM (3 months, 9 days ago)

Quote:

that triangle you drew is how I'd do it



I am genuinely curious why not pick a longer strand, e.g. 3 o'clock?

Thought that you want the fastest strain?

Edited by Yahra (02/05/24 03:44 PM)

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Invisibleelasticaltiger
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Re: Need some agar sectoring suggestions [Re: Yahra] * 1
    #28647784 - 02/05/24 03:38 PM (3 months, 9 days ago)

Quote:

Yahra said:
Quote:

that triangle you drew is how I'd do it



I am genuinely curious why not pick a longer strand, e.g. 3 o'clock?

Though that you want the fastest strain?




I meant the technique of cutting the triangle as he had asked about taking the rope ends.


--------------------
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The Shmuvbox. - The Old TC's Like it :shrug:

Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger!
No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit.

"The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT

Pastywhite's Easy Agar Tek (PastyPlates)

Tiger Drop Video Demos By munchauzen

Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson

EZEKIEL 23:20

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OfflineYahra
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Re: Need some agar sectoring suggestions [Re: elasticaltiger]
    #28647789 - 02/05/24 03:41 PM (3 months, 9 days ago)

Thanks.

:shamecube:

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Invisiblejoze
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Re: Need some agar sectoring suggestions [Re: koma23]
    #28647823 - 02/05/24 03:57 PM (3 months, 9 days ago)

Quote:

koma23 said:
Quote:

joze said:
When you're selecting for isolates and then fruiting them, it's always a gamble what you'll get.




Yeah, I figure as much. But what about if you just want to clone and not isolate, what do you do then with agar?





If want to clone and not isolate (i.e., you want to keep a lot of genetic diversity), all you need to do is make sure you have a clean culture on agar, and then you can send it to grain. You don't need to go through 10 transfers and keep sectoring. You can just transfer it once, make sure it's free of contaminants, and send it. I usually do three transfers from the initial tissue cloning.

Even when you take a transfer from a specific sector, there is going to be a lot of genetic diversity within a teeny tiny sector. You will still see phenotypic variation within your fruits.


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OfflinemorningWood
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Re: Need some agar sectoring suggestions [Re: joze] * 1
    #28647837 - 02/05/24 04:11 PM (3 months, 9 days ago)

That's some really ropey stuff πŸ‘. What strain?!


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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: elasticaltiger]
    #28648340 - 02/06/24 01:40 AM (3 months, 9 days ago)

Quote:

elasticaltiger said:
Very beautiful ropes in those first two pictures.

Personally I would just take a few transfers from the rope ends of either of those first two dishes. Ignore 3rd dish.

If you're hell bent on transfering from the last dish that triangle you drew is how I'd do it.




Yeah, the 3rd and 4th plates are from a different fruit then the first two, so I'm not tossing them, as that fruit was actually better.

Ok, so taking only the end of the ropes (is this how it's officially called?). Appreciate your help mate, tnx!

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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: morningWood]
    #28648341 - 02/06/24 01:42 AM (3 months, 9 days ago)

Quote:

morningWood said:
That's some really ropey stuff πŸ‘. What strain?!




It's all just B+.

Can anybody expand a bit on the ropes? Is it just aggressive rhizo, how much genetic variance is one rope, etc?

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Invisiblemeta_mmxxii

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Re: Need some agar sectoring suggestions [Re: koma23] * 1
    #28648344 - 02/06/24 01:47 AM (3 months, 9 days ago)

Wow those are really impressive plates, I have never seen mycelium structured like that. They almost look fake, though I know they are not. Amazing plates!
I am not the kind of guy that would make art from cult stuff, but that, I would immortalize and frame.
Nice job on those. :rockon:


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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: joze]
    #28648345 - 02/06/24 01:49 AM (3 months, 9 days ago)

Quote:

joze said:
Quote:

koma23 said:
Quote:

joze said:
When you're selecting for isolates and then fruiting them, it's always a gamble what you'll get.




Yeah, I figure as much. But what about if you just want to clone and not isolate, what do you do then with agar?





If want to clone and not isolate (i.e., you want to keep a lot of genetic diversity), all you need to do is make sure you have a clean culture on agar, and then you can send it to grain. You don't need to go through 10 transfers and keep sectoring. You can just transfer it once, make sure it's free of contaminants, and send it. I usually do three transfers from the initial tissue cloning.

Even when you take a transfer from a specific sector, there is going to be a lot of genetic diversity within a teeny tiny sector. You will still see phenotypic variation within your fruits.




Cool. But you still pick a rice grain size patch? And do you see a vastly different fruit with the three transfers? I was thinking of taking basically all of the mycelium and just putting it in LC hehe. I don't quite understand the cloning process as of yet, that is to say, I don't have experience with sectoring and how much that actually changes the end result from the clone ... I know getting a monoculture is the only way to ensure 100% repeatability, but what's the point of cloning a fruit in that scenario if you're changing the genetics via isolating genetics soo much.

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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: meta_mmxxii]
    #28648349 - 02/06/24 01:54 AM (3 months, 9 days ago)

Quote:

meta_mmxxii said:
Wow those are really impressive plates, I have never seen mycelium structured like that. They almost look fake, though I know they are not. Amazing plates!
I am not the kind of guy that would make art from cult stuff, but that, I would immortalize and frame.
Nice job on those. :rockon:




Haha yeah, it's why I'm soo confused about sector selection.

Idk how and why that happens. I used 1.5% wild rice flour and even added in 1% table sugar, so the agar for sure isn't less nutritious so that it would stimulate such rhizo growth.

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Invisiblemeta_mmxxii

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Re: Need some agar sectoring suggestions [Re: koma23]
    #28648352 - 02/06/24 02:00 AM (3 months, 9 days ago)

Quote:

koma23 said:
Quote:

meta_mmxxii said:
Wow those are really impressive plates, I have never seen mycelium structured like that. They almost look fake, though I know they are not. Amazing plates!
I am not the kind of guy that would make art from cult stuff, but that, I would immortalize and frame.
Nice job on those. :rockon:




Haha yeah, it's why I'm soo confused about sector selection.

Idk how and why that happens. I used 1.5% wild rice flour and even added in 1% table sugar, so the agar for sure isn't less nutritious so that it would stimulate such rhizo growth.



Mostly because you are working with a clone. It is very difficult to get that growth from spores or LC. But your plates are impressive none the less.


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OfflineLogicaL ChaosM
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Re: Need some agar sectoring suggestions [Re: koma23]
    #28648365 - 02/06/24 02:11 AM (3 months, 9 days ago)

Trippy mycelium! Looks like an organic snowflake. :eek:

Any ends of the roped-out rhizomorphic mycelium is a good place to sector from.


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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: LogicaL Chaos] * 1
    #28648786 - 02/06/24 10:12 AM (3 months, 9 days ago)

Quote:

LogicaL Chaos said:
Looks like an organic snowflake. :eek:





Ye, exactly my thoughts!

Quote:

LogicaL Chaos said:
Any ends of the roped-out rhizomorphic mycelium is a good place to sector from.




Great, tnx!

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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: meta_mmxxii]
    #28648788 - 02/06/24 10:13 AM (3 months, 9 days ago)

Quote:

meta_mmxxii said:
Mostly because you are working with a clone. It is very difficult to get that growth from spores or LC. But your plates are impressive none the less.




I see. So clones tend to be more ropey, got it.

Edited by koma23 (02/06/24 10:17 AM)

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Invisiblejoze
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Re: Need some agar sectoring suggestions [Re: koma23]
    #28649140 - 02/06/24 03:16 PM (3 months, 8 days ago)

Quote:

koma23 said:
Quote:

joze said:
Quote:

koma23 said:
Quote:

joze said:
When you're selecting for isolates and then fruiting them, it's always a gamble what you'll get.




Yeah, I figure as much. But what about if you just want to clone and not isolate, what do you do then with agar?





If want to clone and not isolate (i.e., you want to keep a lot of genetic diversity), all you need to do is make sure you have a clean culture on agar, and then you can send it to grain. You don't need to go through 10 transfers and keep sectoring. You can just transfer it once, make sure it's free of contaminants, and send it. I usually do three transfers from the initial tissue cloning.

Even when you take a transfer from a specific sector, there is going to be a lot of genetic diversity within a teeny tiny sector. You will still see phenotypic variation within your fruits.




Cool. But you still pick a rice grain size patch? And do you see a vastly different fruit with the three transfers? I was thinking of taking basically all of the mycelium and just putting it in LC hehe. I don't quite understand the cloning process as of yet, that is to say, I don't have experience with sectoring and how much that actually changes the end result from the clone ... I know getting a monoculture is the only way to ensure 100% repeatability, but what's the point of cloning a fruit in that scenario if you're changing the genetics via isolating genetics soo much.



A single mushroom contains multiple genetically distinct organisms coming together to reproduce. It's not just a male and a female. Your clone still contains all the genetic information of all the organisms that came together to form the mushroom. The reason you clone, is because you are eliminating the other genetics in the tub -- you are only cloning the organisms in that one mushroom.

Let's say you have some spores that you germinate, and you fruit them, and you get one really scrawny phenotype, and one stockier phenotype you really like. The scrawny phenotype and the stocky phenotype are going to be different combinations of the genetics from the spores. The stockier phenotype is a genetic mix that you like, so you clone it, to retain those genetics.

Re: your question about a rice size piece vs. a larger piece. For my transfers, I always use a small piece. But for the actual inoculation, I use 1/4 of an agar plate. If you imagine your agar plate as a pizza, I use my scalpel to cut off the outer "crust" leaving only the center. Then I cut the center into quarters, and use those for the inoculation. The reason I cut the "crust" off is because you are more likely to get contaminants along the edges, near the opening of the agar dish.

You could throw in a whole agar dish if you wanted. But I find 1/4 of a dish is plenty to inoculate a quart sized jar of grains, so I can get 4 jars to a plate.


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Offlinekoma23
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Re: Need some agar sectoring suggestions [Re: joze]
    #28650012 - 02/07/24 08:20 AM (3 months, 8 days ago)

Quote:

joze said:
A single mushroom contains multiple genetically distinct organisms coming together to reproduce. It's not just a male and a female. Your clone still contains all the genetic information of all the organisms that came together to form the mushroom. The reason you clone, is because you are eliminating the other genetics in the tub -- you are only cloning the organisms in that one mushroom.

Let's say you have some spores that you germinate, and you fruit them, and you get one really scrawny phenotype, and one stockier phenotype you really like. The scrawny phenotype and the stocky phenotype are going to be different combinations of the genetics from the spores. The stockier phenotype is a genetic mix that you like, so you clone it, to retain those genetics.





Yeah, I totally get that, tnx. I'm unsure about cloning visa vie sectoring. If I select only specific sectors, the resulting fruit wont be a clone anymore ... or is the resulting fruit still in all likelihood going to be 90% the same?

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