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OfflineCameron1996
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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23585423 - 08/27/16 10:45 PM (7 years, 5 months ago)

Maybe change the alkaloid production by isolating on water soluable cbd and thc to help with anticarcinogenic effects and keep cloning.. i was actually going to do this and see how strong i could get it and try it on a sick animal


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Re: Strain Isolations on Agar, Pics and Questions [Re: Cameron1996]
    #23585428 - 08/27/16 10:47 PM (7 years, 5 months ago)

Quote:

Cameron1996 said:
Maybe change the alkaloid production by isolating on water soluable cbd and thc to help with anticarcinogenic effects and keep cloning.. i was actually going to do this and see how strong i could get it and try it on a sick animal




.... What?  Are we still talking about mushrooms here?  Mushrooms getting cancer isn't something we really worry about..  You may come to find that not much in the world can actually be fixed by weed, as nice as that would be from an idealists perspective.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Inocuole]
    #23585447 - 08/27/16 10:57 PM (7 years, 5 months ago)

Quote:

Inocuole said:
It's even more consistent.  You can run actual meaningful tests on it since it is a control at that point.




Ah, so clones can inconsistent results too? I have observed atleast two distinctly different phenotypes in my clone, but chalked it up to growing conditions.


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Re: Strain Isolations on Agar, Pics and Questions [Re: herrenvolk]
    #23585514 - 08/27/16 11:21 PM (7 years, 5 months ago)

Clones can have quite a few sets of genetics present still, yes.


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OfflineCameron1996
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Re: Strain Isolations on Agar, Pics and Questions [Re: Inocuole]
    #23585520 - 08/27/16 11:22 PM (7 years, 5 months ago)

No not the mushrooms getting cancer silly, i ment feed the mushrooms things that fight cancer like water soluable thc and cbd with medical mushrooms like how they can biosynthesize things from different fungi such as ergot stroma tissue with tryptophan and amino acids or how they biosynthesize thc with yeast.. mainly see how growing on a heavy anticarcinogenic chemical content will effect the mycelium and fruit bodys, and clone both so the mushrooms and mycelium dont lose there traits and keep doing that to see if i can get them strong or to produce a whole new chemical in general...try it on pets or animals that are already dieing from cancer and if its successful without the cancer coming back maybe add it to chemo treatments..i have another project i am working on that i dont want to disclose to much because i dont want people trying it and fucking thereselves up but it involves a single spore isolation, do u think i should filter, dilute, then use a glass needle or is there a easyer method? Im going to be working with molds not mycelium at that point


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Re: Strain Isolations on Agar, Pics and Questions [Re: Cameron1996]
    #23585527 - 08/27/16 11:23 PM (7 years, 5 months ago)

So this is on topic... how?  :confused2:


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OfflineTedTheHighlighter
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Re: Strain Isolations on Agar, Pics and Questions [Re: Cameron1996]
    #23585538 - 08/27/16 11:27 PM (7 years, 5 months ago)

Okay, I understand how most monocultures will be weaker than a choice clone. You can visibly see the best mushroom to clone but it takes luck and testing to find the best monoculture after months of isolating.
So what are the chances you get that monoculture that yields consistant, excellent grows that are even better than grows from a nice clone?
Just wondering how beneficial isolating actually is


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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23585547 - 08/27/16 11:30 PM (7 years, 5 months ago)

About x%.  :shrug:  The chance is there, but there's no cut and dry way to say for sure how long you could spend isolating cultures before you find something that makes it worthwhile.


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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23585554 - 08/27/16 11:32 PM (7 years, 5 months ago)

The chances of getting a good isolate with good yield and potency? Uh I highly doubt anyone will have an answer. Also depends on what you call good yield and potency. For my standards, I'd say I'd be looking at 1 in 50 isolates would be what I want.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Mad Season]
    #23585588 - 08/27/16 11:47 PM (7 years, 5 months ago)

My definition of good enough would be better than any clone you could take.
Simoly because, while isolating a strain is an awesome thing, I don't see how it is worth the months of work if you can just clone and get better results


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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23585619 - 08/28/16 12:08 AM (7 years, 5 months ago)

And that's why I've always done clones. Isolating is something I'd do if you have a lot of spare time to go genetics hunting


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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23586052 - 08/28/16 07:53 AM (7 years, 5 months ago)

Quote:

TedTheHighlighter said:
My definition of good enough would be better than any clone you could take.
Simoly because, while isolating a strain is an awesome thing, I don't see how it is worth the months of work if you can just clone and get better results




Isolates like that exist, but that's not to say another clone couldn't crop up in the future that's better still.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Inocuole]
    #23586451 - 08/28/16 10:37 AM (7 years, 5 months ago)

Soooooo

Say you isolate 100 strains and find the ideal one. Is that ideal strain likely to be a better producer than an ideal clone from a MS grow?

Basically, is an awesome monoculture better than an awesome clone?


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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23586608 - 08/28/16 11:36 AM (7 years, 5 months ago)

It could be, until you find a better clone, and then that could be the best until you find a better isolate.  There is no ceiling of how "awesome" something can be, so, without knowing what measurements you're using to gauge awesomeness, it's hard to say.  All that anyone could say for sure is that they'll be different in some way.

If you want something more consistent than a clone, which will sometimes do different things, then you isolate it further.  If you don't mind a little bit of variability, you stick with clones.

It's all far too random for anyone to deifinitively answer any questions like that about it though, because things change, life keeps mutating and crossing and doing its own thing.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Inocuole]
    #23586684 - 08/28/16 12:03 PM (7 years, 5 months ago)

Yeah I understand.
I was just wondering because cloning is much simpler than isolating so I figured that isolating will eventually give you your best possible grows. Otherwise, it seems not worth it when you can just clone and get the same results


--------------------
Alice asked the Cheshire Cat, who was sitting in a tree, “What road do I take?”

The cat asked, “Where do you want to go?”

“I don’t know,” Alice answered.

“Then,” said the cat, “it really doesn’t matter, does it?”


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Re: Strain Isolations on Agar, Pics and Questions [Re: TedTheHighlighter]
    #23586807 - 08/28/16 12:40 PM (7 years, 5 months ago)

Overall I don't recommend isolating unless you have a goal in mind that requires it. :shrug:  But there are merits and everybody should probably do it at least once.


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Re: Strain Isolations on Agar, Pics and Questions [Re: Inocuole]
    #23587224 - 08/28/16 03:04 PM (7 years, 5 months ago)

I'm currently doing transfers from MS plates, hoping to clean the mycelium of bacterial contaminations from the dirty spore print. But I'm wondering if this process is also isolating "strain groups", strains that go well together and run intertwined on the agar plate. The cultures were transferred 3 times. (I did the agar way too soft for the 2 first transfers).

I've made some bottle/v-tek to test my subcultures (3 replicates for each of the 8 partial isolates) and maybe get some clones. The results should be a bit more consistent than straight MS jars, right ?

Maybe I should have tried some MS BRF cakes, fruit the possibly contaminated cakes, then clone ? What are the advantages of going the agar way, beside learning the technique ?


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Re: Strain Isolations on Agar, Pics and Questions [Re: Cameron1996]
    #23587310 - 08/28/16 03:32 PM (7 years, 5 months ago)

Quote:

Cameron1996 said:
No not the mushrooms getting cancer silly, i ment feed the mushrooms things that fight cancer like water soluable thc and cbd with medical mushrooms like how they can biosynthesize things from different fungi such as ergot stroma tissue with tryptophan and amino acids or how they biosynthesize thc with yeast.. mainly see how growing on a heavy anticarcinogenic chemical content will effect the mycelium and fruit bodys, and clone both so the mushrooms and mycelium dont lose there traits and keep doing that to see if i can get them strong or to produce a whole new chemical in general...try it on pets or animals that are already dieing from cancer and if its successful without the cancer coming back maybe add it to chemo treatments..i have another project i am working on that i dont want to disclose to much because i dont want people trying it and fucking thereselves up but it involves a single spore isolation, do u think i should filter, dilute, then use a glass needle or is there a easyer method? Im going to be working with molds not mycelium at that point




Very confusing/confused post my friend.

1. Neither THC nor CBD are water soluble, at all. That's the whole principle behind most hash making procedures

2. THC and even CBD don't really have anti carcinogenic effects, that is a misunderstanding. THC and CBD are often used to treat the nausea and anorexia associated with cancer, certainly not anti carcinogenic though

3. The compounds that have shown some potential for actually treating certain types of cancer are not the psychoactive compounds, but rather the 270ish other nonpsychoactive compounds found in cannabis, with more being discovered all the time

4. Lol genetic engineering, a la splicing the ability to produce spider silk into goats, etc, is a LOT more complicated than that. Every time it has been done it involved a whole team of PhDs with gov grants (nearly infinite resources and brain power), and geneticists competing for the Nobel prize. There is some amazing potential in engineering molds to produce hard-to-manufacture compounds, but it would require a few genetic engineering degrees to even scratch the surface on

5. "Single spore isolation" is not a thing (well, not beyond monokaryotic isolates ). Spore reproduction processes basically are analogous to sexual reproduction, with genetic material from 2 spores. I highly doubt you want a monokaryotic isolate, I think you are confused

6. Mold is still mycelium, just a different species. Like how you and a naked mole rat are different species, but both have blood

Read Paul Stamets books TMC and Mycelium Running, should clear up a lot and will fill your brain with wonder :smile:

Quote:

TedTheHighlighter said:
Yeah I understand.
I was just wondering because cloning is much simpler than isolating so I figured that isolating will eventually give you your best possible grows. Otherwise, it seems not worth it when you can just clone and get the same results




The "same", definitely not. It's similar, but the distinguishing feature lies in that an isolate is a single strain, and clones usually have multiple strains present. It depends what you are trying to do and how you define "good". If you are just trying to grow some good mushrooms, isolates are altogether unnecessary and clones are much preferable. But if your intention is to run meaningful tests or have cultures you could bet on, isolates are essential

Mad and inocuole, great info, very helpful for all involved :highfive1:

Quote:

Piezo said:
I'm currently doing transfers from MS plates, hoping to clean the mycelium of bacterial contaminations from the dirty spore print. But I'm wondering if this process is also isolating "strain groups", strains that go well together and run intertwined on the agar plate. The cultures were transferred 3 times. (I did the agar way too soft for the 2 first transfers).

I've made some bottle/v-tek to test my subcultures (3 replicates for each of the 8 partial isolates) and maybe get some clones. The results should be a bit more consistent than straight MS jars, right ?

Maybe I should have tried some MS BRF cakes, fruit the possibly contaminated cakes, then clone ? What are the advantages of going the agar way, beside learning the technique ?




1. These strain groups you are referring to are fascinating to me as well, I have had a couple of cultures which were perfectly round for the first 10 transfers, only started showing sectors on the 11th. These groups are good in that they mean compatible strains that can network together, but a pain in the ass trying to isolate

2. Partial isolate isn't a thing, it's either an isolate or not. What you're referring to is what many call "limited MS" because the herd of strains has been thinned from the initial MS. More consistent? Not necessarily, because you never know which strain(s) will become most established or how well they will play with the other strains present.

3. Don't fool with contaminated cakes, waste of time. Figure out where the failure was and fix it. You say the spore print was dirty, so probably here.  And most importantly, "the agar way" is not an alternative to "learning the technique "; agar is no substitute for technique. If you have bad technique on agar it will get old fast. Agar requires even more technique than no agar

4. There are many uses for agar: doing clones, isolations, cleaning cultures, etc. just like any other tool or technique, the advantage lies in how you employ it, not the tool itself. In your case, agar is perfect for cleaning up a dirty spore print. You will want to take transfers from the leading edge of the myc until you are sure you have a clean culture, only then would you inoculate


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Edited by c10h12n2o (08/28/16 03:59 PM)


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Offlinespacechildo
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Re: Strain Isolations on Agar, Pics and Questions [Re: c10h12n2o]
    #23587358 - 08/28/16 03:46 PM (7 years, 5 months ago)

:firstladyofapproval:


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Re: Strain Isolations on Agar, Pics and Questions [Re: c10h12n2o]
    #23587619 - 08/28/16 05:04 PM (7 years, 5 months ago)

Thanks for the reply.
Quote:

c10h12n2o said:
1. These strain groups you are referring to are fascinating to me as well, I have had a couple of cultures which were perfectly round for the first 10 transfers, only started showing sectors on the 11th. These groups are good in that they mean compatible strains that can network together, but a pain in the ass trying to isolate

2. Partial isolate isn't a thing, it's either an isolate or not. What you're referring to is what many call "limited MS" because the herd of strains has been thinned from the initial MS. More consistent? Not necessarily, because you never know which strain(s) will become most established or how well they will play with the other strains present.

3. Don't fool with contaminated cakes, waste of time. Figure out where the failure was and fix it. You say the spore print was dirty, so probably here.  And most importantly, "the agar way" is not an alternative to "learning the technique "; agar is no substitute for technique. If you have bad technique on agar it will get old fast. Agar requires even more technique than no agar

4. There are many uses for agar: doing clones, isolations, cleaning cultures, etc. just like any other tool or technique, the advantage lies in how you employ it, not the tool itself. In your case, agar is perfect for cleaning up a dirty spore print. You will want to take transfers from the leading edge of the myc until you are sure you have a clean culture, only then would you inoculate





2. Yeah, by "partial isolate" I was referring to those "strain groups" obtained from limited MS. If I understand correctly, the replicated jars from the same limited MS plate could give different results because the strain demographics might diverge in each jar ?

3. The print was made in a hurry in a septic and humid environment... I haven't tried to inoculate a jar with it yet.
That's why i thought it would be a good time to learn agar work. By "learning the technique" I was referring to the agar work. Like you said in 4, I thought this situation would be the perfect time to learn it.

I think my agar plates are clean now (I will take pictures during the next transfer). However, I'm worried about my genetic choices. They must be supported by test runs and that's a lot of work and time to be spent...

The alternative I was mentioning is to let the nature to do its natural selection thing in the jar inoculated with MS, and then clone the best looking fruit body. It seems to be much less work...


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