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InvisibleMycomedicine888
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
    #28498771 - 10/10/23 12:27 PM (3 months, 16 days ago)

So this seems to work really well for cubes and gourmet types, but would it do so for the faster scenescing stuff like Pans or Cordyceps too?


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I may have inhaled some of the spores....



Copelandia cyanescens 2023


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Offlinetree frog
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Re: The OFFICIAL Stasis Storage Thread [Re: Mycomedicine888]
    #28501275 - 10/12/23 07:37 AM (3 months, 14 days ago)

I've been wanting to try the colonized grain method.

"Submerge colonized grain in sterile water and store in the fridge.  Water can be replaced with sterilized mineral oil to protect cold weather sensitive species such as paddy straw." - Organic Mushroom Farming and Mycoremediation, Tradd Cotter

Basically fish a stasised grain out of storage and inoculate a plate or lc with it.

I've used agar wedges in sterile water to store pink oysters too.  Seemed to work well but I only kept them a year.


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Listen to the silence behind the engines' noise.  Jesus, Sweets, listen.  Hear it?  It's a love song.
For whom?
You are loved.
~ David Foster Wallace, Westward the Course of Empire Takes Its Way


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OfflineDERRAYLD
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Re: The OFFICIAL Stasis Storage Thread [Re: tree frog]
    #28501279 - 10/12/23 07:40 AM (3 months, 14 days ago)

You can just as easily store dried grain vs myceliated grain in a suspension.


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Offlinetree frog
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Re: The OFFICIAL Stasis Storage Thread [Re: DERRAYLD]
    #28501333 - 10/12/23 08:31 AM (3 months, 14 days ago)

Quote:

DERRAYLD said:
You can just as easily store dried grain vs myceliated grain in a suspension.




I looked at faht's post, linked on the first page.  Drying grains seems like I'd have to build new lids for.

Is there an easier tek for getting the grains to dry?  Maybe shake a few out into the bottom of a sterilized half pint jar and cap with my current GE lids?


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Listen to the silence behind the engines' noise.  Jesus, Sweets, listen.  Hear it?  It's a love song.
For whom?
You are loved.
~ David Foster Wallace, Westward the Course of Empire Takes Its Way


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OfflinesopposS
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
    #28508358 - 10/17/23 04:16 PM (3 months, 9 days ago)

saveed


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Offlinebongoman
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Re: The OFFICIAL Stasis Storage Thread [Re: soppos]
    #28509677 - 10/18/23 04:15 PM (3 months, 8 days ago)

I'm exploring agar wedges in distilled H2O as my main and preferred approach to archiving cultures.

One remaining question I have is whether the final transfer plate should be water agar so there's no nutrition in the agar wedge being put into stasis?

Thoughts on this?


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InvisibleReverendMyc

Registered: 03/29/19
Posts: 1,494
Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
    #28509685 - 10/18/23 04:25 PM (3 months, 8 days ago)

I use regular nutritious agar wedges. Haven't done side by side testing, but figure that there is no need to add the additional stress of starvation to a culture I am trying to preserve.

The op mentions using wood or paper as a dense nutrient source but :shrug:

If someone has better information please share it.


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LAGM 2.024
Stoned Gummys tek (Gummies from sclerotia or mushrooms) *Not just for stones any more
How to succeed in mycology (and life) - know nothing, read everything, try something, and accept advice.
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Offlinebongoman
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Re: The OFFICIAL Stasis Storage Thread [Re: ReverendMyc]
    #28509693 - 10/18/23 04:34 PM (3 months, 8 days ago)

Thanks. Regular agar would be much simpler.

This paper infers they are just using regular nutritious agar, or at least it doesn't specifically mention water agar.

https://sci-hub.st/10.1139/w08-049


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OfflineNichrome
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman] * 3
    #28509851 - 10/18/23 06:47 PM (3 months, 8 days ago)

I store lc in sealed vials for years on end. It's not exactly the same as a wedge in water but is similar conceptually. I have a Cordyceps that still kicks off like it should from just one drop from the banked vial. Same with semperviva, tamp, etc. I'd make a solid edumacated guess that a nutritious agar wedge in water would work just fine and there is no need for special agar prep for such a storage method.

I've cut pieces of stem and kept them in baggies for 6 months before getting around to plating them as clones. Mycelium, circumstantially, is capable of extreme longevity.


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Better to be deprived of food for three days, than tea for one.


Freedom is not the right to do as you please, but the liberty to do as you should. ~Emerson


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Offlinebongoman
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Re: The OFFICIAL Stasis Storage Thread [Re: Nichrome]
    #28513421 - 10/21/23 07:39 PM (3 months, 5 days ago)

Quote:

Nichrome said:
I store lc in sealed vials for years on end.




Are you using slants as well? I'm trying to decide whether I can use this method as my sole form of archiving cultures.


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InvisibleMycolorado
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
    #28513424 - 10/21/23 07:41 PM (3 months, 5 days ago)

Dry grain spawn FTW.


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OfflineSerMantis
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
    #28519899 - 10/27/23 08:46 AM (2 months, 30 days ago)

Quote:

bongoman said:
I'm exploring agar wedges in distilled H2O as my main and preferred approach to archiving cultures.

One remaining question I have is whether the final transfer plate should be water agar so there's no nutrition in the agar wedge being put into stasis?

Thoughts on this?




I've seen woodies (Lion's mane) fruit off of a toothpick in a water stasis vial. It'll probably go to stasis after it eats the full pick. If the point of stasis is to preserve a specific culture, that fruit would be putting spores into the liquid meaning there is now more genetic variance. That is better than cloning fruit back to agar after every grow and good enough for a lot of use cases, but maybe not for multi year (or decade) long preservation of specific genetics.

I'd think that you are right with water agar or the SFD. It would make sense to limit nutrients in the solution, but I haven't made any stasis tubes like this yet to try it.


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Wood is good.


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InvisibleFloret
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Re: The OFFICIAL Stasis Storage Thread [Re: SerMantis]
    #28566151 - 12/03/23 12:24 AM (1 month, 25 days ago)

These long term experiments are great :popcorn:


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LAGM 2024


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OfflineGuyOnaBuffalo
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Re: The OFFICIAL Stasis Storage Thread [Re: SerMantis]
    #28566624 - 12/03/23 11:28 AM (1 month, 24 days ago)

I don't think it would eat the pick. Once it's in an environment lacking free oxygen and all present oxygen is bound to hydrogen, it should go right into stasis.


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Offlinec01h
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Re: The OFFICIAL Stasis Storage Thread [Re: GuyOnaBuffalo]
    #28585835 - 12/16/23 06:34 PM (1 month, 11 days ago)

:threadmonitor:


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Offline3.A.M
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Re: The OFFICIAL Stasis Storage Thread [Re: c01h]
    #28605424 - 01/01/24 05:28 AM (26 days, 19 hours ago)

Without me going all the way back through this thread, do we know if dried grains have an average life span or can they be stored indefinitely?

And if anyone’s interested I’ve been using small autoclave pouches with great success for storing and drying grains, also use them for my swabs, they’re sanitary and literally designed to wick away moisture and dry out while at the same time maintaining a protective shield against contamination, I just pour grains straight from the jar in them when I do my g2g process, seal them and let them dry from inside the pouch, super cheap and almost too easy.


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Offlinec01h
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Re: The OFFICIAL Stasis Storage Thread [Re: 3.A.M]
    #28605605 - 01/01/24 09:42 AM (26 days, 15 hours ago)

at least 2+ years based of this post


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Offline3.A.M
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Re: The OFFICIAL Stasis Storage Thread [Re: c01h]
    #28605841 - 01/01/24 01:09 PM (26 days, 11 hours ago)

Sweet!


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Invisiblethe_chosen_one
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Re: The OFFICIAL Stasis Storage Thread [Re: 3.A.M] * 4
    #28609668 - 01/04/24 04:40 PM (23 days, 8 hours ago)

How is it a person can be here for years and all the sudden something like this pops up and it was started a year ago? :confused:

I've become a huge fan of the 2ml cryovials and the Castellani method.


OR

Just forget some syringes in the fridge for 13 years. :lol: We'll see these in a decade or so.


Works for spores though. :shrug: I've labelled these Tamp 13 for just that reason lol.


Thanks to Tree Frog for the link!
:manofapproval:


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"Luck favors the observant." - Workman



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OfflineSeventhMushroom
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Re: The OFFICIAL Stasis Storage Thread [Re: the_chosen_one] * 1
    #28609792 - 01/04/24 06:23 PM (23 days, 6 hours ago)

Just adding my experience here too:

I used 20ml glass vials  using 10ml of a low-nutrient solution (0.1% lme + tap water) to store several cultures. This was in April of last year, so 8+ months now.

(this kind, with rubber stoppers, coincidentally the exact same stoppers used for SHIPs on jar lids.)


As far as I can tell, these worked amazing for "stasis"! I even did a simple experiment when I first started using these vials:

I filled 3 vials with a different amount of liquid. (5ml, 10ml, and 15ml) Then added a drop of LC, and watched what happened. The 5ml of liquid had the *most* growth, with the 15ml vial stopping growth much earlier. This would make sense if the oxygen is the limiting factor. I went with 10ml for the rest of my work though, as that's just enough to fill a 10ml syringe! wow! :laugh:

Last week I got to see my first confirmation that it worked! I took two samples out (lions mane, and some blue oysters) and put some on plates. No fancy "revival" procedure, just opened the vial, used a syringe to drop some on the plate. The plates are doing great, quickly colonized, no contamination (that I can tell). *and* the liquid left in the vial started growing thicker again!! After that confirmation, I went ahead and dumped the rest of the vials into some grain jars a few days ago, they are also doing great.


All this to say, I think using wooden sticks, or paper, and stressing about a nutrient-deprived medium is over complicated and unnecessary. See you in 5 years to confirm ;P


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LAGM 2024


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