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InvisibleAshtray161
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Registered: 03/21/21
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Why Slants Instead of Agar Plates?
    #27397371 - 07/22/21 12:42 AM (2 years, 6 months ago)

Hello all!
So im starting to finally get the grasp of things enough that im starting to chase down and trying to preserve my best cultures from my best results of my first few grows so I can start to get these wonderful canopies I see consistently and am starting to look into how exactly to go about that.
Im starting to look into slants and masters and trying to figure it all out and Im curious what the advantage is of storing a culture on a slant rather than just on an agar petri dish?
Also I see that cloning from a clone can start to degrade the genetics, how quickly does that happen/how many generations? Is the best thing to clone, take spores, grow from spores, clone again, repeat?


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Invisiblestubb
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Re: Why Slants Instead of Agar Plates? [Re: Ashtray161]
    #27397441 - 07/22/21 02:51 AM (2 years, 6 months ago)

Mainly because they won't dry out as fast as a plate.  They seal better, there's less air volume in the tube than the plate, and there's more moisture locked in thicker agar.

I don't have an answer to your second question. 

Quote:

Is the best thing to clone, take spores, grow from spores, clone again, repeat?



Depends what you're aiming to do...  :shrug:  You can also just keep growing from a slant culture if you want.


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OfflineDouglas Fir
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Registered: 08/04/21
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Re: Why Slants Instead of Agar Plates? [Re: stubb]
    #27418235 - 08/07/21 01:17 PM (2 years, 5 months ago)

From my understanding, the slant is for storage of the culture. You could hypothetically transfer over to fresh agar over and over again for the rest of time, but if you would like to save a culture without doing that; you would make a slant with distilled water and some wood and then -  perhaps refrigerate?


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Invisiblenosf3r4tu


Registered: 03/26/19
Posts: 775
Re: Why Slants Instead of Agar Plates? [Re: Douglas Fir]
    #27418246 - 08/07/21 01:28 PM (2 years, 5 months ago)

Nice to see you here Douglas. You first post! Haha
Anyways...
The best way to preserve cultures is distilled, sterile water storage. That's 10% LC and 90% distilled sterilized water. It can be stored at room temperature for 2 - 5 years.


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OfflineDouglas Fir
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Re: Why Slants Instead of Agar Plates? [Re: nosf3r4tu]
    #27418584 - 08/07/21 04:49 PM (2 years, 5 months ago)

lol I have been here for a while, just made a new account because 19 year old me was a little to flamboyant with information about myself. However, slants were something I was researching recently so I thought I would contribute.


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Invisiblepaperbackwriter
Edward Lear


Registered: 03/31/14
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Re: Why Slants Instead of Agar Plates? [Re: Douglas Fir]
    #27419165 - 08/08/21 06:07 AM (2 years, 5 months ago)

Quote:

Douglas Fir said:
From my understanding, the slant is for storage of the culture. You could hypothetically transfer over to fresh agar over and over again for the rest of time, but if you would like to save a culture without doing that; you would make a slant with distilled water and some wood and then -  perhaps refrigerate?




It is for culture storage.  The problem with just transferring on plates is senescence (aging).  All cells can only divide so many times before they lose vigor and die.

We keep cultures on slants because we can basically slow down the metabolism of young cells by either denying it essential nutrients (the sterile water tek just mentioned which I use for warm weather species like pink oyster, tamps, and king tuber) or by tossing it in the fridge.  In the later case you want to make sure the culture has enough food and moisture to survive its dormancy.  The slant gives both.  The stick provides complex nutrients.  The slant itself makes sure there's enough moisture because it's a lot of agar in a space without much surface area.

Then, when we want to grow out some mushrooms, we're always pulling young and vigorous cells.


--------------------
Why should we strive with cynic frown
To knock their fairy castles down?  ~ Eliza Cook

It's rather embarrassing to have given one's entire life to pondering the human predicament and to find that in the end one has little more to say than, 'Try to be a little kinder.' ~Aldous Huxley


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Invisiblethe_chosen_one
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Re: Why Slants Instead of Agar Plates? [Re: paperbackwriter]
    #27420093 - 08/08/21 05:20 PM (2 years, 5 months ago)

Exactly. Even the best sealed petri dish will dry out eventually in a refrigerator. Slants will too, but it's typically many years before it does.


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InvisibleAshtray161
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Registered: 03/21/21
Posts: 4,503
Loc: Rugby, England
Re: Why Slants Instead of Agar Plates? [Re: the_chosen_one]
    #27420103 - 08/08/21 05:29 PM (2 years, 5 months ago)

Thanks for the clarification all :smile:
ive picked up some sticks lol


--------------------
(You Know What Time It Is)
Major Issues in the Psychedelic Movement: https://www.shroomery.org/forums/showflat.php/Number/27677086:elmo:
"You never have to prove the fool a fool, just let them speak."
Please, be an adult. Get vaccinated. Dont use psychedelics as an excuse. Dont come at me with some hippy dippy nonsense, GO GET VACCINATED.
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Invisiblec10h12n2o
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Registered: 01/21/15
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Re: Why Slants Instead of Agar Plates? [Re: Ashtray161]
    #27421857 - 08/09/21 10:04 PM (2 years, 5 months ago)

i absolutely love slants. so many advantages over long term plates

- as mentioned, they dont dry out fast like plates do. i think the oldest plates ive used were like 1-1.5 yrs old, where i have used 6 yr old slants without any issue

- i always make slants at full strength standard MEA (3%) rather than the mycostandard 1.5%, since there is no real reason to try to make the myc grow out and sector or search for nutes

- after you take some myc from a slant w a loop, there is fresh agar underneath, in the damaged area that the existing myc will colonize

- soooo much better for storage space optimization. you can fit way more slants in the same fridge space

- i make about 100 slants at a time and just leave them out in room temp in a ziploc till i need them. i do the same with plates, but the slants last longer and dont dry out

- great workflow for testing new cultures or strain hunting. when you want to test a new  culture, just slant it, apply a unique identifier (i use stickers from a big roll of number stickers 0000-9999, just stick one on the slant), then label all grows/spawn/etc with that numerical ID. then when you grow them out, update your notes for each culture based on that numerical ID, and you can pull the slant out of the fridge anytime you want to revive it

as far as senescence (your second question) this is kinda up in the air. its considered best practice to avoid unneeded expansion (ie instead of multiplying the same grain master 40x, every so often go back to the original master slant). the same concerns are why people avoid cloning clones. some people claim to see senescence after just a few generations, but personally i dont think i have ever seen it... and it makes me wonder about if it ever happens at all in nature, for example with that Humungous Fungus in oregon which is over 3.5 sq miles and estimated to be over 8000 yrs old

but i think instead of "is it ok to clone a clone?" a better question would be "why would you clone a clone?" maybe it makes sense if you are playing with the different sectors you will find in most clones, but if you are happy with a culture and you already have the same culture slanted or on a master dish/jar, why not just go back to the master?


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Offlinesporemaster420
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Registered: 02/11/23
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Re: Why Slants Instead of Agar Plates? [Re: nosf3r4tu]
    #28197865 - 02/21/23 06:03 PM (10 months, 30 days ago)

Quote:

nosf3r4tu said:
Nice to see you here Douglas. You first post! Haha
Anyways...
The best way to preserve cultures is distilled, sterile water storage. That's 10% LC and 90% distilled sterilized water. It can be stored at room temperature for 2 - 5 years.




You can do this? Isn’t the distilled water a bit hypotonic for the mycelium?


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OfflineAlan RockefellerM
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Re: Why Slants Instead of Agar Plates? [Re: sporemaster420] * 1
    #28204347 - 02/25/23 03:58 PM (10 months, 26 days ago)

Quote:

sporemaster420 said:
Quote:

nosf3r4tu said:
Nice to see you here Douglas. You first post! Haha
Anyways...
The best way to preserve cultures is distilled, sterile water storage. That's 10% LC and 90% distilled sterilized water. It can be stored at room temperature for 2 - 5 years.




You can do this? Isn’t the distilled water a bit hypotonic for the mycelium?





My distilled water tubes lasted a lot longer than my slants did.  A few years later most of my slants didn't grow and most of the distilled water tubes did.  They were stored at room temperature though I imagine they would last even longer in the fridge.

I took a colonized plate and cut it up into little squares, then put those into sterile distilled water in 50 ml centrifuge tubes.


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OfflineEclipse3130
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Re: Why Slants Instead of Agar Plates? [Re: Alan Rockefeller]
    #28206184 - 02/26/23 10:26 PM (10 months, 24 days ago)

That's good to know. My experience it depends on the genetic how long it lasts in a slant specifically. Some die out and don't recover as well as others.


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