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OfflineRyeJar
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Registered: 04/16/15
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Keeping track of age
    #21898796 - 07/04/15 10:06 PM (8 years, 6 months ago)

I understand the p value system but what I don't understand is when and what to assign a value

I grew spores out on grain, then grew it out on substrate, grew out fruits, grew out tissue samples on agar, transfered 4 times so far at the size of a dime or so (so not the whole plate)

Now when I do get a few clean isolates Staments in GGMM says to label those p0 or p1 (I forgot and the book is packed away)

RR in his vids doesnt cover it in detail if I remember correctly, but at one point it seems he infers that your first transfer from fruit to plate starts the 'aging' and needs to be tracked

It makes sense to me to start tracking age at spore germination though.  Why would we not?  There are a lot of cell divisions going on before tissue ever even hits agar! (mostly in the jar depending on your shaking habits I assume) And then 10 transfers later we start tracking?

I would assume it's hard to track cell divisions when not in a petri although so it seems only fair to start tracking as soon as it hits a plate contrary to the book GGMM


Edited by RyeJar (07/04/15 11:39 PM)


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OfflineRescueShroom
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Re: Keeping track of age [Re: RyeJar]
    #21898902 - 07/04/15 10:30 PM (8 years, 6 months ago)

i dont have the im monitoring this post picture... but.. im monitoring this post...


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OfflineMachiavelliavore
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Re: Keeping track of age [Re: RescueShroom]
    #21899437 - 07/05/15 01:29 AM (8 years, 6 months ago)

The main idea as I understand it is to track the volume of expansion of a given sample.  A couple agar transfers won't degrade it much effectively.  In MS, this will be hard to track, because we don't know how much of the volume in composed of a single strain.

In her culturing tek, violet suggests starting with tiny invitro grain substrate (1/8th cup,) and harvesting ideal looking pins to get the youngest source for your master culture, which will have expanded very little, compared to something expanded to 5qt grain, then another 11 of coir, then a fatty mushroom.

Seems like a good idea to me.  Can't say I know the answer to your notation question.  I call my master X, then if I grow that out into a master jar for an upcoming spawn run, I call that jar X', and then after I G2G that Jar, I call those other jars X''.  If for some reason my master had already been spawned out, I'd leave it as X' or X'' or whatever, then add additional ' to denote further expansion.

An invitro grain sub can probably be estimated as almost zero expansion, as can agar plates.


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


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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OfflineRyeJar
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Re: Keeping track of age [Re: Machiavelliavore]
    #21900664 - 07/05/15 11:54 AM (8 years, 6 months ago)

Quote:

Machiavelliavore said:

In her culturing tek, violet suggests starting with tiny invitro grain substrate (1/8th cup,) and harvesting ideal looking pins to get the youngest source for your master culture, which will have expanded very little, compared to something expanded to 5qt grain, then another 11 of coir, then a fatty mushroom.






That's a good idea

Quote:

Machiavelliavore said:
An invitro grain sub can probably be estimated as almost zero expansion, as can agar plates.




Agar plates can't be counted as 0 expansion I'm pretty sure.  The basis of the p value comes from the size of a mycelial cell and the size of the petri (usually 100 mm) to keep track of expansion I believe.


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InvisibleBuck513

Registered: 04/17/14
Posts: 5,682
Re: Keeping track of age [Re: RyeJar]
    #21900701 - 07/05/15 12:04 PM (8 years, 6 months ago)

You can make plenty of a2a transfers.

You can only expand a culture 2 or 3 times in jars because you keep shaking them, weakening the mycelium. You are obviously not shaking the mycelium when doing a2a so that allows for more transfers.


--------------------
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OfflineRyeJar
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Re: Keeping track of age [Re: Buck513]
    #21901138 - 07/05/15 01:45 PM (8 years, 6 months ago)

Quote:

Buck513 said:
You can make plenty of a2a transfers.

You can only expand a culture 2 or 3 times in jars because you keep shaking them, weakening the mycelium. You are obviously not shaking the mycelium when doing a2a so that allows for more transfers.




It s3ems like shaking would produce less cell divisions since you are distributing the mycelium throughout the jar making it have to travel less to fully colonize


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InvisiblePastywhyteMDiscord
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Re: Keeping track of age [Re: Buck513]
    #21901206 - 07/05/15 02:01 PM (8 years, 6 months ago)

Quote:

Buck513 said:
You can make plenty of a2a transfers.

You can only expand a culture 2 or 3 times in jars because you keep shaking them, weakening the mycelium. You are obviously not shaking the mycelium when doing a2a so that allows for more transfers.




You can still G2G a few dozen times before even a hint of slowing down will show. Keep your masters young but you can expand the hell outta it once its outta the slant.


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InvisibleBuck513

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Re: Keeping track of age [Re: Pastywhyte]
    #21901472 - 07/05/15 03:04 PM (8 years, 6 months ago)

Quote:

Pastywhyte said:
Quote:

Buck513 said:
You can make plenty of a2a transfers.

You can only expand a culture 2 or 3 times in jars because you keep shaking them, weakening the mycelium. You are obviously not shaking the mycelium when doing a2a so that allows for more transfers.




You can still G2G a few dozen times before even a hint of slowing down will show. Keep your masters young but you can expand the hell outta it once its outta the slant.



I've never heard you could expand it a few dozen times. But I'll take your word for it. That seems rather crazy though lol


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InvisiblebodhisattaMDiscordReddit
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Re: Keeping track of age [Re: Buck513]
    #21901500 - 07/05/15 03:11 PM (8 years, 6 months ago)

you can expand until the wheels fall off of a culture but it's more than dozens of g2g generations. it's all about how soon on you do the expansions. temporal limitations are much more important than the expansion itself. you can expand the fuck out of mycelium but you want to do it soon that's why go back to masters or slants either way though if you do g2g as soon as jars are ready you could do it 20 times in a row and still have a good culture.


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OfflineRyeJar
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Re: Keeping track of age [Re: bodhisatta]
    #21901585 - 07/05/15 03:35 PM (8 years, 6 months ago)

Quote:

bodhisatta said:
Temporal limitations are much more important than the expansion itself.




What do you mean by this?

Also, when and what do we designate as p0 then?


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InvisiblebodhisattaMDiscordReddit
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Re: Keeping track of age [Re: RyeJar]
    #21901603 - 07/05/15 03:39 PM (8 years, 6 months ago)

if you do 20 g2g generations in 2 months it's way better than doing 20 g2g generations over 6 months.


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OfflineRyeJar
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Re: Keeping track of age [Re: bodhisatta]
    #21902759 - 07/05/15 08:43 PM (8 years, 6 months ago)

Gotcha.  So should time elapsed in vegatative state start being used alongside the p value system for a more accurate description of the cultures age?


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Re: Keeping track of age [Re: RyeJar]
    #21902810 - 07/05/15 08:54 PM (8 years, 6 months ago)

likely


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Invisibleazur
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Re: Keeping track of age [Re: Pastywhyte]
    #21903309 - 07/05/15 11:04 PM (8 years, 6 months ago)

Quote:

Pastywhyte said:
Quote:

Buck513 said:
You can make plenty of a2a transfers.

You can only expand a culture 2 or 3 times in jars because you keep shaking them, weakening the mycelium. You are obviously not shaking the mycelium when doing a2a so that allows for more transfers.




You can still G2G a few dozen times before even a hint of slowing down will show. Keep your masters young but you can expand the hell outta it once its outta the slant.



A2A will eventually weaken out. Yes, myc will still grow out. But fruiting will lack. Trust me. I've done it.


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


A cube is NOT a cube.

FALL IN LOVE WITH LC
FOTTSE!!!
ALL NOOBS READ THIS!!!



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InvisiblePastywhyteMDiscord
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Re: Keeping track of age [Re: azur]
    #21903315 - 07/05/15 11:06 PM (8 years, 6 months ago)

What is A2A?


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Invisibleazur
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Re: Keeping track of age [Re: Pastywhyte]
    #21903324 - 07/05/15 11:07 PM (8 years, 6 months ago)

That's what buck said. Agar 2 agar


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


A cube is NOT a cube.

FALL IN LOVE WITH LC
FOTTSE!!!
ALL NOOBS READ THIS!!!



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InvisiblePastywhyteMDiscord
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Re: Keeping track of age [Re: azur]
    #21903359 - 07/05/15 11:16 PM (8 years, 6 months ago)

I just call it a transfer. It can eventually weaken after a huge amount but it would take a long time. But no one keeps their masters in room temperature, slant em,  the fridge saves all.


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Invisibleazur
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Re: Keeping track of age [Re: Pastywhyte]
    #21903368 - 07/05/15 11:17 PM (8 years, 6 months ago)

I just call it transfer as well. Was trying to stay in tune with this thread.
I have a culture i recently ran weak from  way too many transfers


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


A cube is NOT a cube.

FALL IN LOVE WITH LC
FOTTSE!!!
ALL NOOBS READ THIS!!!



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InvisibleBuck513

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Re: Keeping track of age [Re: azur]
    #21903379 - 07/05/15 11:20 PM (8 years, 6 months ago)

How many is too many


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Invisibleazur
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Re: Keeping track of age [Re: Buck513]
    #21903385 - 07/05/15 11:20 PM (8 years, 6 months ago)

A lot!  Several months worth


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


A cube is NOT a cube.

FALL IN LOVE WITH LC
FOTTSE!!!
ALL NOOBS READ THIS!!!



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InvisiblePastywhyteMDiscord
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Re: Keeping track of age [Re: azur]
    #21903399 - 07/05/15 11:23 PM (8 years, 6 months ago)

Bummer. I usually try to keep em at a minimum before slanting then I don't really give a crap how treat the working culture.  Beat it up and toss when done. Usually I only expand a few times before i feel like a change but I do have a clone I have been riding for over a year now. Its a workhorse.


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Invisibleazur
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Re: Keeping track of age [Re: Pastywhyte]
    #21903421 - 07/05/15 11:27 PM (8 years, 6 months ago)

Yea. Mine shit the bucket after 9 months


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


A cube is NOT a cube.

FALL IN LOVE WITH LC
FOTTSE!!!
ALL NOOBS READ THIS!!!



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InvisiblePastywhyteMDiscord
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Re: Keeping track of age [Re: azur]
    #21903473 - 07/05/15 11:37 PM (8 years, 6 months ago)

That one is done, definitely weaker. But was G2G and transferred so many times I lost count. It was a beast. Its gonna be another year before I bother to revive it.


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Invisibleazur
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Re: Keeping track of age [Re: Pastywhyte]
    #21904128 - 07/06/15 06:06 AM (8 years, 6 months ago)

Nice. Good to know about g2ging so far down the line


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


A cube is NOT a cube.

FALL IN LOVE WITH LC
FOTTSE!!!
ALL NOOBS READ THIS!!!



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