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Offlinepsilocyanide
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What is a generation
    #24871642 - 12/27/17 02:17 PM (7 years, 21 days ago)

What exactly "counts" as gaining a generation?

Like, if I transfer (agar/grain/LC) to (agar/grain/LC)?  I am asking because I understand that you arent supposed to go past a certain number of generations because the mycelium loses its power.

I recently asked this question and bodhisatta (thanks for answer) said:

Quote:

bodhisatta said:
1. Yes. Shouldn't be a problem sterilized. Try it out and let us Know though.

2. I think you're seeing the same things but it never "disappears" in the LC after a "shake"

3. Spores to fruit to spores is a gen. Other expansion can be called a generation but i would call them expansions.

4. Ya know when you know




But this answer doesnt make sense to me.  I always thought that you cant do g2g too many times, and also it seems like starting with spores is like starting fresh.

So pretty much, i know (or think, atleast), that I cant g2g too many times, but can I transfer LC to LC or LC to grain, or agar to agar, etc. only a limited number of times?


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OfflinehamloafM
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Re: What is a generation [Re: psilocyanide]
    #24871649 - 12/27/17 02:20 PM (7 years, 21 days ago)

Spores to fruit is a generation.  Starting with spores is starting fresh.  You can't do g2g as many times as you like because you lose p-value, and encourage the senescence of the culture.


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Offlinepsilocyanide
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Re: What is a generation [Re: hamloaf]
    #24871663 - 12/27/17 02:30 PM (7 years, 21 days ago)

Ok, so I am confusing the word then for something else.  Thanks. But can I also not (lc/agar)2(lc/agar), or grain2(agar/LC) or (agar/LC)2grain?

If im not being clear, do you lose p-value from any transfer, or does this only have to do with grain?


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InvisibleMorelman
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Re: What is a generation [Re: psilocyanide] * 1
    #24871922 - 12/27/17 04:54 PM (7 years, 21 days ago)

P values are given to transfers of mycelium from one medium to the next.  First transfer being P1 and each transfer after that being given a +1. It's very easy to rack up P values when chasing isolates of mycelium. Increased P values (transfers) leads to senescence.
Like Ham said, a generation (F1,F2, etc.) is spore to fruit. The spores from those fruits create the next generation.
By P4, I have my ms cultures spawned to a bulk substrate; P3 if I'm working with a clone. The first two or three transfers are monitored for contaminants.


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InvisiblebodhisattaMDiscordReddit
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Re: What is a generation [Re: psilocyanide] * 1
    #24872025 - 12/27/17 05:46 PM (7 years, 21 days ago)

P value is an absolutely stupid name given to the term expansion by paul stamets. Who wanted to name something else after himself.

G2g isn't a generation. But its expansion.

Paul's p value also doesn't take into consideration the age of your cultures/spawn.

If you take fully colonized jars let then sit a week then do g2g is less optimal than taking freshly colonized jars and doing g2g.

Basically the p value scale is retarded shit. Hence why only one mycologist uses it. Time spent out of cold storage is arguably more important than the amount of expansion done to a culture.

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InvisiblebodhisattaMDiscordReddit
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Re: What is a generation [Re: bodhisatta]
    #24872027 - 12/27/17 05:48 PM (7 years, 21 days ago)


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Offlinepsilocyanide
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Re: What is a generation [Re: bodhisatta]
    #24872381 - 12/27/17 08:41 PM (7 years, 21 days ago)

Thanks.  Very helpful.

This quote from the search you left also cleared up any questions I had.

Quote:

RogerRabbit said:
Quote:

ozzysmygod said:
Its not about how many 'generations', more about how much the mycelium has traveled/spread




This is correct.  Most of you are using the word 'generation' wrong in this thread.  A generation is going from spores to fruits and back to spores again for the next generation.

When it comes to grain to grain transfers, there's not a set 'number' of transfers you can do.  Obviously, 5 grain to grain transfers with pints is a bit different than 5 grain to grain transfers with large spawn bags.  Age of the mycelium also plays a role.  If you let the grains sit for a week or two after colonization before G2G, you can't do as many transfers before senescence sets in.

When I'm rapidly expanding mycelium, I'll do 5 G2G transfers before calling it good and starting over with a fresh, low P-value culture.
RR




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InvisibleMateja
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Re: What is a generation [Re: Morelman]
    #24872530 - 12/27/17 10:12 PM (7 years, 21 days ago)

Quote:

Morelman said:
Increased P values (transfers) leads to senescence.




Just from transferring? Are you sure? I though senescence was something rare and that one really had to take it to stupid levels to get to that point?


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InvisibleMorelman
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Re: What is a generation [Re: Mateja]
    #24872715 - 12/28/17 03:44 AM (7 years, 21 days ago)

Like any organism, cells can only divide so many times before becoming senescent.


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Edited by Morelman (12/28/17 08:10 PM)

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OfflinePeacefrog
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Re: What is a generation [Re: Morelman]
    #24872723 - 12/28/17 04:16 AM (7 years, 21 days ago)

Yes from my understanding and experience with it, cell division is where senescence comes in to play. One can transfer MANY times from agar to agar with very minimal cell divisions. When isolating down for a mono culture from spores, one could end up transferring and growing out 20-100 times really, without loosing any vigor.  I have several slanted cultures that are a several years old and have been transferred so many times I have lost count.

They still grow exactly the same way they did when I isolated and tested them. My favorite Cope/pan culture (in my signature) is 7-8 years old now and has been rejuvenated many times (I.e. transferred from a slant to a new agar plate, then back to a fresh slant) and it is still “young” mycelium and grows very well.

If one were to take that mycelium to several G2G transfers and spawn pounds of substrate, a ton of cell divisions will have taken place in order to colonize all of that. A2A transfers doesn’t require many cell divisions IME.

My 2 pennies.


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InvisibleMorelman
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Re: What is a generation [Re: Peacefrog]
    #24872845 - 12/28/17 07:16 AM (7 years, 20 days ago)

A combination of refrigeration and/or lack of nutrients slows down cell division.


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OfflineShroominmojo0213
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Re: What is a generation [Re: Morelman]
    #24872887 - 12/28/17 08:06 AM (7 years, 20 days ago)

I had read or heard that you can master a clone out up to ten times (Or a grain jar,etc.) I master some out up to 5 times but that's it. Than I start over from a spore,Agar,l.i.) I concur on a generation as being a spore to a fruiting body though.

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Re: What is a generation [Re: Morelman]
    #24872891 - 12/28/17 08:09 AM (7 years, 20 days ago)

Your thinking and work has proved to be correct in my experience as well! Your working towards getting the best bang for your buck by transferring plates.

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Re: What is a generation [Re: Morelman]
    #24873017 - 12/28/17 09:09 AM (7 years, 20 days ago)

Quote:

Morelman said:
Like any organism, cells can only divide so many times before becoming senescencent.



That's not true. Some organisms have no senescence or negligible senescence

https://en.m.wikipedia.org/wiki/Negligible_senescence

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Re: What is a generation [Re: bodhisatta]
    #24873165 - 12/28/17 10:15 AM (7 years, 20 days ago)

Does this relate to why it has been advised to not take a clone from a grow that was cultured from a clone itself?


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Re: What is a generation [Re: Morelman]
    #24873407 - 12/28/17 11:47 AM (7 years, 20 days ago)

Does anyone has documents that show how cube mycelium looks and performs after too many transfers?


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Re: What is a generation [Re: Germs]
    #24873776 - 12/28/17 02:47 PM (7 years, 20 days ago)

Quote:

Germs said:
Does this relate to why it has been advised to not take a clone from a grow that was cultured from a clone itself?




:thumbup:


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Re: What is a generation [Re: Germs]
    #24873794 - 12/28/17 02:51 PM (7 years, 20 days ago)

Quote:

Germs said:
Does this relate to why it has been advised to not take a clone from a grow that was cultured from a clone itself?



Cloning from a clone is a mop job from poor handling


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OfflinePeacefrog
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Re: What is a generation [Re: cronicr]
    #24874676 - 12/28/17 08:28 PM (7 years, 20 days ago)

@Mateah
I’ve personally never experienced senescence, so I do not have any pictures. What I can show is a slanted mono culture after ~6-8 months of isolation and countless transfers:

After all those A2A transfers, it was still vigorous and fast to colonize grain and fruit. Sadly, I no longer have that culture. I’m not a cube grower anymore and was only illustrating isolation techniques on another site for members who had not started agar yet, or may be intimidated by agar work. So I only tested it once in a mini grow and it was mediocre:


But it went through many plates of agar before I tossed it without showing any sign of senescence or aging.

My pan culture (mentioned above) has been A2A transferred probably ~30-40 times and still grows very well and is super fast to colonize and fruit:


Morelman is absolutely correct when he says a combo of refrigeration and/or a lack of nutrition helps slow/stop cell division. There are a lot of great posts around the OMC about using plain distilled water for long term storage. I have never tried it personally, as I just like using slants in the ref., but I’m sure it’s a very solid method from my research.

As far as cloning from a clone goes, I would say it works but why would one do it unless it’s a last ditch effort to save a coveted culture. But I’m a proponent of isolating down a clone to test if any strains are better than the combined genetic profile of the mixed MS fruit body. Sometimes you can isolate a gem. So there doesn’t seem to be too much difference from cloning a clone except the extra cell division it would take to colonize a substrate and make the fruit body itself.

I have read this discussed many times and I guess I just never wanted to test the boundaries of a culture that much.


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Re: What is a generation [Re: Peacefrog]
    #24874691 - 12/28/17 08:36 PM (7 years, 20 days ago)

Wauw thank you! :smile:

Im saving this post!
Also you just gave me an idea to experiment with :cheers:


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