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OfflineSkizor1337
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LC to Agar to Print?
    #27357958 - 06/21/21 06:03 PM (2 years, 7 months ago)

I'm just seeking a little clarification

Say you take a multispore syringe
Create LC and then inoculate jars, spawn them, and from the new fruits make a print

Now with that print you make a spore syringe

With that you go to Agar.

Take the agar and transfer it to LC

Now with that (isolated LC if you will) is it safe to say any fruit you produce with that is all the same "strain"

OR after using the newly created LC and making fruits to make new prints from that to further isolate the culture?


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OfflineHindsight
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Re: LC to Agar to Print? [Re: Skizor1337]
    #27358118 - 06/21/21 08:12 PM (2 years, 7 months ago)

Phase 1:
Say you take a multispore syringe
Create LC and then inoculate jars, spawn them, and from the new fruits make a print


Phase 2:
Now with that print you make a spore syringe
With that you go to Agar.
Take the agar and transfer it to LC


I'm just trying to be sure I understand what you are asking.

First question for you is: Is there any reason in Phase 1 you go MS syringe to LC but then in phase 2 you go MS syringe to agar?

Second question is: When you say go to agar, do you mean just to ensure you have a clean culture to put to grain or do you mean take many transfers on agar to try to isolate the mycelium down to what one would call an isolate?


Without having answers to those question, I'm guessing you're asking how best to create spore prints that, when grown out, produce fruit characteristics exactly as you want as reliably and consistently as possible? If so, agar vs LC doesn't matter. You just need to repeat the grow from MS inoculation -> Fruit -> Spore Print From Most Desired Fruit -> Grow From MS Inoculation of that Spore Print -> Repeat until you are getting consistent results in your fruits.


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Offlineherbnmyco
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Re: LC to Agar to Print? [Re: Hindsight]
    #27358217 - 06/21/21 09:29 PM (2 years, 7 months ago)

If you want an isolated culture make a clone on agar. Take a fruit you like, rip the stem in half and scrape a tiny piece from the inside. Try not to touch anything but the inside of the fruit. Then put that little piece on agar and it will turn back into mycelium. Also ditch the syringe its just an extra step that can introduce contams unless thats how you got spores.


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OfflineSkizor1337
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Re: LC to Agar to Print? [Re: Hindsight]
    #27358268 - 06/21/21 10:24 PM (2 years, 7 months ago)

Quote:

Hindsight said:
Phase 1:
Say you take a multispore syringe
Create LC and then inoculate jars, spawn them, and from the new fruits make a print


Phase 2:
Now with that print you make a spore syringe
With that you go to Agar.
Take the agar and transfer it to LC


I'm just trying to be sure I understand what you are asking.

First question for you is: Is there any reason in Phase 1 you go MS syringe to LC but then in phase 2 you go MS syringe to agar?

Second question is: When you say go to agar, do you mean just to ensure you have a clean culture to put to grain or do you mean take many transfers on agar to try to isolate the mycelium down to what one would call an isolate?


Without having answers to those question, I'm guessing you're asking how best to create spore prints that, when grown out, produce fruit characteristics exactly as you want as reliably and consistently as possible? If so, agar vs LC doesn't matter. You just need to repeat the grow from MS inoculation -> Fruit -> Spore Print From Most Desired Fruit -> Grow From MS Inoculation of that Spore Print -> Repeat until you are getting consistent results in your fruits.




I have a read a lot but just trying to wrap my head around it all.

The MS syringe I started with was just from a vendor

Aren't the spores from a single print isolated in a way?

And I go to agar to ensure clean culture Yes.

Just didn't know if a print used in LC/Agar is technically the same as cloning to agar.

I figured spore print to agar would be better since you can create a ton of streak plates compared to just cloning with a single fruit.

Unless it's subjective but trying to dial in my grows a bit more.


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OfflineHindsight
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Re: LC to Agar to Print? [Re: Skizor1337]
    #27358495 - 06/22/21 06:02 AM (2 years, 7 months ago)

The spores from a single print are nowhere near as isolated as what you get from growing out a clone from a tissue sample of a fruit. A mushroom has a lot of genetics that it drops in its spores, and when two spores  germinate and two compatible hyphae join up and “mate”, it’s like when two humans get together….. there are a lot of genes and some dominant and recessive traits and because of this, you just don’t know for sure what you’ll end up with. Odds are good it will be similar to what the parent mushroom was but you can still see variations.

A spore print to LC/Agar is not the same as cloning because with cloning, you aren’t changing the genetics involved. With cloning, you know that the exact same combination of mycelium and their current genetics that all worked together to make that fruit will be exactly the same when you grow it out again (or it should be - there are some unexplained weirdnesses on occasion). With a clone, you don’t have the genetic shuffling that a mushroom fruit goes through when producing spores nor do you have the randomness introduced when spores germinate into hyphae and then join with another hyphae to “mate” and become mycelium that grows out from there. And when you do an MS grow, you likely have TONS of different mycelium cultures all growing at once and competing or cooperating. You could have 3 working together to fruit while also having ten that won’t fruit and are just taking up resources. So if you grow out a clone, you’re eliminating all the other mycelium that was competing and may have been responsible for producing small or weird fruit or no fruit at all.

When cloning, you can create as many plates as you want. Or better yet, clone to agar and then transfer that to LC and use the LC to inoculate as many spawn jars as you like. I personally think LC is better for clone perpetuation than taking many transfers from agar because I have seen people inadvertently isolate their clones on agar by taking transfers of just one part of the mycelium and that then having the unintended consequence of creating a mycelium colony that’s missing something the original clone had and as a result, it won’t fruit or work the same.


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OfflineSkizor1337
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Re: LC to Agar to Print? [Re: herbnmyco]
    #27440880 - 08/24/21 11:27 AM (2 years, 5 months ago)

Quote:

herbnmyco said:
If you want an isolated culture make a clone on agar. Take a fruit you like, rip the stem in half and scrape a tiny piece from the inside. Try not to touch anything but the inside of the fruit. Then put that little piece on agar and it will turn back into mycelium. Also ditch the syringe its just an extra step that can introduce contams unless thats how you got spores.




I got the cloning down it's from that point where I'm not sure where to go

Like take the clone agar and transfer to agar? Or transfer the agar to grain which is a slow process from previous trials.

I'm trying to keep the isolate going that's why I considered using LC because of how much you get


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OfflineA.k.aM
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Re: LC to Agar to Print? [Re: Skizor1337]
    #27440973 - 08/24/21 12:21 PM (2 years, 5 months ago)

Anytime you use spores it’s like a genetic reset.


Taking a piece of agar and making lc with it is just like putting that same piece onto a new plate, only now it’s in liquid form instead of agar.


Usually what people do is take a piece of mushroom to clone and put it on a plate, then grow it out and transfer a couple times. From there you should get pretty consistent results so you spawn a tub with it and see what you get. If it’s good then you can keep the plate around as a master.


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OfflineSkizor1337
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Re: LC to Agar to Print? [Re: A.k.a]
    #27445413 - 08/27/21 04:58 PM (2 years, 5 months ago)

Quote:

A.k.a said:
Anytime you use spores it’s like a genetic reset.


Taking a piece of agar and making lc with it is just like putting that same piece onto a new plate, only now it’s in liquid form instead of agar.


Usually what people do is take a piece of mushroom to clone and put it on a plate, then grow it out and transfer a couple times. From there you should get pretty consistent results so you spawn a tub with it and see what you get. If it’s good then you can keep the plate around as a master.





Thanks! It's been a while since I made any


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