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fossilshark
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The OFFICIAL Stasis Storage Thread 21
#28161262 - 01/28/23 12:06 PM (11 months, 23 days ago) |
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I've been waiting a long time to make this thread as this is something I am very interested in. The goal of this thread is to share the many different methods of storing cultures in stasis. I will be posting my method and I would love for everyone to contribute their method of storing cultures in stasis. I in NO way came up with stasis storage and it has been around in many different forms for decades.
What is stasis storage? Stasis storage is storing a sterile sample of mycelium in a no-oxygen no-nutrient environment causing the mycelium to go into stasis ceasing all metabolic activity.
Why stasis storage? There are multiple advantages of stasis storage over other long term culture storage methods.
#1: The biggest advantage of stasis storage is the complete inaction of metabolic activity. Other long term culture storage methods (slants, cryo) only slow the metabolic activity down. In theory this can lead to changes in the culture that is not desired. Furthermore this means that unlike slants you do not have to "wake up" the mycelium at any point to continue to preserve it. Stasis has been shown to be effective for 30 years in https://sci-hub.st/10.1139/w08-049 and thats just when they stopped the study. In theory stasis storage is indefinite.
#2: Stasis storage is super simple and can be done with solid media or liquid or just straight up agar wedges. Very universal technique for anyone's style.
#3: My favorite part, stasis storage works at room temperature. Absolutely no need to take up valuable space in your moms fridge. 
My method:
My initial trials 2 years ago were done to preserve Enigma x Rusty Whyte F2 from Solipsis using paper strips. I plan on switching over to wood skewers to make things easier to handle. First I hydrated paper strips by dunking paper strips in distilled water for a few seconds and sterilizing them for 20 minutes in my trusty presto along with some vials of distilled water.
After the paper strips and water were prepped I laid the strips on colonized germ plates (more strains more better baby) and let the mycelium colonize them for about a week.

After the paper strips were colonized I transferred them into vials of distilled water (along with some fat plate pins to try and clone them in the future).

And done. Thats the whole tek. Stasis storage is stupid simple. I ran the cultures about a year after I made them and they bounced back very quickly and I plan on doing a 2 year test next month when I have time.
NOTES: While the original stasis vials I made are useable its apparent that my sterile technique back then was sub par. One of the vials had a scoby on top but the mycelium was just fine.
For coprophilic species paper strips, wood skewers, toothpicks ECT work just fine with no issues. I theorize that if you want to store lignophilic species this would be too much available nutrients and effectively make the solution into LC. This would still work as once LC runs out of nutrients it goes into stasis, but to me that does not seem ideal. A better solution would be to use SFD discs like in the original paper.
I appreciate any questions or discourse on this thread on the topic. I think this method is an amazing tool for a mycology enthusiast. I appreciate those of you who take the time to read through this post.
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VietboyX
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28161273 - 01/28/23 12:12 PM (11 months, 23 days ago) |
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Nice write up, will definitely be helpful to link to the nubs in discord for long time storage
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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: VietboyX]
#28161281 - 01/28/23 12:16 PM (11 months, 23 days ago) |
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Great write up, been waiting for this!
What are you sterilizing the strips in after soaking, just any pp5 container?
Would using wood skewers still use the same process... soak to hydrate, pc, lay on a colonized plate, then into sterile distilled water?
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fossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye]
#28161293 - 01/28/23 12:23 PM (11 months, 23 days ago) |
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I use pyrex test tubes from kartersci. Yes skewers would work exactly the same and I will be switching over to those for future cultures since they are easier to handle then paper strips.
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Nextcontestant
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28161310 - 01/28/23 12:34 PM (11 months, 23 days ago) |
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fahtster
Now With 33%More Faht



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Re: The OFFICIAL Stasis Storage Thread [Re: Nextcontestant] 4
#28161534 - 01/28/23 03:33 PM (11 months, 23 days ago) |
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Lids for drying out colonized grains for dry stasis
Ppl have also just left some kernels in the bottom of their qts (after securing the lid back on) and let them dry out that way but I hear it can take quite a while.
I’m going to doing a lot of drying of cultures out in the future and probably simplify my lid set up so it’s just a small SHIP and use only two layers of tyvek with a widemouth 1/2 pint.. putting that in a bag of desiccant (damprid) with the larger lid and less barriers should speed things up by another week I’m thinking. Right now I’m at about 3 weeks from hydrated to dry where I can store as is or load into swab packs to take up less space
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bakedbeings
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
#28161548 - 01/28/23 03:45 PM (11 months, 23 days ago) |
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fossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
#28161690 - 01/28/23 05:17 PM (11 months, 23 days ago) |
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This method is really interesting, so you think the mycelium goes into stasis despite the presence of oxygen? So basically it isn't exclusively no-oxygen no-nutrient but can also just be caused by a lack of water?
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Pastywhyte
Say hello to my little friend



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 2
#28162481 - 01/29/23 08:01 AM (11 months, 22 days ago) |
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Interesting thread here. Gives me some ideas for when I’m gonna need to back up my library. Nice contribution
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bakedbeings
orbiter of truth


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Re: The OFFICIAL Stasis Storage Thread [Re: Pastywhyte]
#28162493 - 01/29/23 08:13 AM (11 months, 22 days ago) |
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from what ive read so far it seems like i should be able to just take a clean tissue sample and stick it in some water, no?
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fahtster
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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings] 2
#28162509 - 01/29/23 08:32 AM (11 months, 22 days ago) |
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Quote:
fossilshark said: This method is really interesting, so you think the mycelium goes into stasis despite the presence of oxygen? So basically it isn't exclusively no-oxygen no-nutrient but can also just be caused by a lack of water?
Well there’s no visible growth after many months.. if there is cell division, it’s happening at a speed that isn’t noticeable after a certain point of dryness. I just revived this mak 118 jar that’s looked exactly like this since the end of February


I think if you remove any part of those three, oxygen, food, water, you can get stasis or at least a degree of it.
That mak 118 is just about start knotting/pinning so we’ll see how it does. You’d think after being dried out for 10 months it’d slow it down or wear out. We shall see. Ime, it does not

I actually already revived the clone once this season using a separate dried out jar but I was trying to laymix in a bag… didn’t work out so well and I had to remove the block from the container so fruit out the side pins.

So I kept it normal this time around as far as sub construction and I think it’ll be even better
But I also revived this fp+ culture that was dried for 8 months
Before..

After revival..
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fossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings]
#28162739 - 01/29/23 11:36 AM (11 months, 22 days ago) |
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Baked that is correct. If you get a sterile sample of tissue you can just put it in sterile water and boom that clone is saved for generations to come.
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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
#28162740 - 01/29/23 11:37 AM (11 months, 22 days ago) |
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Wow super dope. I might save some cultures like this in the future. I am very excited to run those gandalf grains you sent me a while back.
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bakedbeings
orbiter of truth


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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28162757 - 01/29/23 11:44 AM (11 months, 22 days ago) |
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Quote:
fossilshark said: Baked that is correct. If you get a sterile sample of tissue you can just put it in sterile water and boom that clone is saved for generations to come.
sweet
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dna24
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28162774 - 01/29/23 11:54 AM (11 months, 22 days ago) |
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i have been super interested in this since i first read about long term storage and currently am testing this myself. i am at nowhere the time frame to see if im doing it correctly yet but thats half the fun. i need to invest in some quality vials.
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fossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: dna24] 1
#28163355 - 01/29/23 06:45 PM (11 months, 22 days ago) |
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Thats why this thread is 2 years in the making. I wanted to make sure it works before I started posting about it.
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fungusul
Fungus Kingdom


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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28164960 - 01/30/23 08:02 PM (11 months, 21 days ago) |
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Thanks for sharing!
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Range Spacer
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28166204 - 01/31/23 04:22 PM (11 months, 20 days ago) |
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Am I correct in my assumption that one cannot put liquid spores into stasis and that it would be better just to keep a spore print?
Edited by Range Spacer (01/31/23 04:24 PM)
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HILLBILLY OUTLAW
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28166219 - 01/31/23 04:37 PM (11 months, 20 days ago) |
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--------------------
 🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 TEAM SPREAD THE LOVE! Smellyhobbit said: Embarrassment and bashfulness are leeches on your ability to learn.
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fossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: Range Spacer]
#28167021 - 02/01/23 03:31 AM (11 months, 19 days ago) |
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Technically speaking (I may be wrong about this) because spores by themselves with no nutrient will not reproduce they are already in stasis.
As far as I know spore syringes probably wouldn't stay viable for infinitely long periods of time even with proper storage, implying that it is either not true stasis or there is other factors that prevent spores from germinating after X amount of time.
Why not just germinate those spores and put the resulting culture into stasis?
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Guerrilla
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28167030 - 02/01/23 03:54 AM (11 months, 19 days ago) |
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-------------------- Being pissed on does not make you a real man.
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AyePlus
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Re: The OFFICIAL Stasis Storage Thread [Re: Guerrilla]
#28167111 - 02/01/23 06:00 AM (11 months, 19 days ago) |
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Hate losing cold storage cultures. Definitely trying this out
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LadysKnight
Hello Ladies


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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28167113 - 02/01/23 06:03 AM (11 months, 19 days ago) |
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Quote:
fossilshark said:
After the paper strips and water were prepped I laid the strips on colonized germ plates (more strains more better baby) and let the mycelium colonize them for about a week.
Since spores are dirty, germ plates are dirty. I guess this is where you got the contam.
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fossilshark
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Re: The OFFICIAL Stasis Storage Thread [Re: LadysKnight]
#28167217 - 02/01/23 08:24 AM (11 months, 19 days ago) |
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Probably lol. I definitely didnt lose the genetics tho as ive been able to revive them on 2 seperate occasions.
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towndaze
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28168599 - 02/02/23 06:33 AM (11 months, 18 days ago) |
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Does that mean we finally have a mass-market ready Rusty Whyte & APE distribution technique?
Because shit son with this you could create [url=https://www.shroomery.org/forums/showflat.php/Number/27331873#27331873 ]giga mega super ultra isolated genetics across thousands of sheets of paper[/url] worth of completely standardized mycelium. Is it natural? Of course not. Is it useful? Quite likely.
-------------------- Time is nature's way of making sure that everything doesn't happen at once.
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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: towndaze]
#28169710 - 02/02/23 07:30 PM (11 months, 18 days ago) |
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Not a clue what you just said but hell yes brother I love the energy.
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nektar61
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28173923 - 02/05/23 07:50 PM (11 months, 15 days ago) |
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Fascinating stuff. Thank you.
I'm curious why this will grow and old spores won't seem to. I know someone revitalized a 20 plus year old print for Redboy, but I've seen people fail to grow spores that are under 5 years old. I believe stasis works but don't know why it does and storing spores seems to not work well.
Would removing all oxygen help? What about storing cultures without the water, and in argon or similar? I know that's complicating something simple, but I'm wondering if a tiny bit of oxygen is needed.
I looked up that arctic seed vault where Bill Gates stores his seeds to grow on all the farm land he's buying after he and his friends wreck the world (just kidding, maybe), this is the tek. It's seeds, not growing culture, and plants, not fungus, but interesting:
"The seeds are stored in sealed three-ply foil packages and then placed into plastic tote containers on metal shelving racks.[51] The storage rooms are kept at −18 °C (−0.4 °F). The low temperature and limited access to oxygen will ensure low metabolic activity and delay seed ageing. The permafrost surrounding the facility will help maintain the low temperature of the seeds if the electricity supply fails."
https://en.wikipedia.org/wiki/Svalbard_Global_Seed_Vault#Seed_storage
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B Traven
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Re: The OFFICIAL Stasis Storage Thread [Re: nektar61]
#28173969 - 02/05/23 08:32 PM (11 months, 15 days ago) |
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-------------------- Beware of advice- even this.
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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: nektar61] 3
#28174161 - 02/05/23 09:53 PM (11 months, 15 days ago) |
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Spores are just gametes and not fully developed cells so they wouldnt have the resources to completely shut down and stay viable.
Based on what methods people use at the moment if you get rid of just one of the three oxygen, water, or nutrients the mycelium will go into stasis and be viable until it is either destroyed or wakes up.
Stasis storage of mycelium cultures needs no overcomplicating.
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towndaze
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28174227 - 02/05/23 10:35 PM (11 months, 15 days ago) |
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Quote:
fossilshark said: Not a clue what you just said but hell yes brother I love the energy.
Sorry I was high. And now I'm high again. Let's break it down. Many more exotic variants don't or don't easily produce spores, which heavily restricts their spread. If you do a super isolated LC then freeze it onto paper you can create hundreds of hyper-standardized vials a day.
-------------------- Time is nature's way of making sure that everything doesn't happen at once.
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Sulfurshelfsean
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Re: The OFFICIAL Stasis Storage Thread [Re: towndaze]
#28174393 - 02/06/23 05:09 AM (11 months, 14 days ago) |
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I've had lcs last years. One out out clean mycellium for six years but eventually they contam. I always use very little nute so I always wondered if they were in stasis. After about a year the culture liquid is completely clear. This looks like a much better way to store thank you for the write up. I'm going to try this.
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Hindsight
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Interesting idea. I do the same thing but just take a piece of colonized agar and put it in sterile distilled water vial. It has worked well for me. There is, of course, a small amount of nutrient from the agar, but it doesn't seem to impact storage life. Paper is a nutrient source so it seems to be a roughly equal approach.
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bakedbeings
orbiter of truth


Registered: 09/01/20
Posts: 4,218
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Quote:
Sulfurshelfsean said: I've had lcs last years. One out out clean mycellium for six years but eventually they contam. I always use very little nute so I always wondered if they were in stasis. After about a year the culture liquid is completely clear. This looks like a much better way to store thank you for the write up. I'm going to try this.
ive noticed the same thing with low nute LCs. they colonize about 80% and then freeze
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side-eye
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Re: The OFFICIAL Stasis Storage Thread [Re: bakedbeings] 4
#28181951 - 02/11/23 08:50 AM (11 months, 9 days ago) |
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I dig it so I tried it.
I got some 5mL tubes with a rack, and wood stir sticks. Each one breaks into three or four pieces that fit in the tubes. Then I filled a bunch of the bottles with distilled water.


I soaked the stir sticks in the bottle for about 30 minutes, figured they needed a little more time than paper. Poured water out of the stir stick bottles, then PC'd everything in my IP for 45 minutes. Stir sticks onto a few plates. These pictures were taken two days apart, myc is already starting to get into the wood. Be sure all the broken pieces are ripped apart from each other, you can see how the Reishi plate has a big V shape that I didn't want to risk pulling apart.
 
Once this grows into the wood I'll drop em into the distilled water. I'll report back in 30 years to check if your tek actually works.
-------------------- LAGM 2023
Edited by side-eye (02/11/23 08:51 AM)
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nektar61
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye] 3
#28182942 - 02/11/23 07:32 PM (11 months, 9 days ago) |
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Quote:
side-eye said: I'll report back in 30 years to check if your tek actually works.
Excellent.
I have an experiment I've set up on here that should last 15 years. (vacuum packing very dry clone shrooms into 2 gram packs, then taking one every year and reporting back. First one will be pretty soon.
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fahtster
Now With 33%More Faht



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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster] 4
#28182962 - 02/11/23 07:45 PM (11 months, 9 days ago) |
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Quote:
fahtster said: That mak 118 is just about start knotting/pinning so we’ll see how it does. You’d think after being dried out for 10 months it’d slow it down or wear out. We shall see. Ime, it does not

Bag did aight.

The question is if it’s able to go 10 months dried out at room temp and come back and do that ^^^ what’s stopping it from going 10 more, or even more than that?
I have dried out cultures in jars that are over a year old. I can revisit this thread in a couple years and give those cultures a try at reviving.. see ya then
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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster] 1
#28190459 - 02/16/23 04:09 PM (11 months, 4 days ago) |
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1
Alright so I got two plates' sticks in water.

I ran into a few problems on this one. First was I let the plates colonize the sticks a little too much, especially on the Reishi plate. I tried picking it up with tweezers, but it was so stuck to the plate that I lifted up the whole puck of agar out of the dish. I tried cutting it out but it still grabbed any agar underneath the stick. Reishi was by far the worst, but all of the sticks lifted a bunch of myc off the plate. Here's how colonized the plates were:

Second, the wood sticks float. These tubes are 5mL so pretty short, and when I let go of the stick it floated about halfway out of the tube. I used the lid to push them down as I capped em. Longer tubes/sticks may not be as bad, and paper may not be as bad either. I'm worried they picked up some bac hitting the rim of the tube, so I will give this another go soon with paper vs sticks.
Lastly, my fault, I dropped some sticks in the wrong tube of water. Not sterile so I had to toss those. Don't work high, kids.
-------------------- LAGM 2023
Edited by side-eye (02/16/23 04:10 PM)
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Guerrilla
Bumbaclart


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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye] 4
#28190466 - 02/16/23 04:17 PM (11 months, 4 days ago) |
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What's the purpose of using wood or paper when a wedge of myc will suffice and last indefinitely? Seems like you're making it way more complicated than it needs to be.
-------------------- Being pissed on does not make you a real man.
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Kabbal
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Re: The OFFICIAL Stasis Storage Thread [Re: Guerrilla]
#28199567 - 02/22/23 05:15 PM (10 months, 29 days ago) |
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Good shit here Boyo. I will post some results from 5 months ago when you told me about it.
-------------------- For noobs starting out READ THIS: README Journal 2.0 (on going) Grow Bag Experiment(on going) Liner/Mesh Experiment(on going)
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mycodependent
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28221141 - 03/08/23 05:08 PM (10 months, 15 days ago) |
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oh fuck yeah I'm trying the fuck outta this shit ASAP.
-------------------- We have been inoculated and are spreading spores LAGM 2.023 TATs

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174BPM



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Re: The OFFICIAL Stasis Storage Thread [Re: mycodependent]
#28248914 - 03/27/23 08:32 AM (9 months, 27 days ago) |
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astra
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28254070 - 03/30/23 04:25 AM (9 months, 24 days ago) |
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dna24
Darth Randal



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 1
#28254230 - 03/30/23 07:53 AM (9 months, 24 days ago) |
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Quote:
fossilshark said: Spores are just gametes and not fully developed cells so they wouldnt have the resources to completely shut down and stay viable.
Based on what methods people use at the moment if you get rid of just one of the three oxygen, water, or nutrients the mycelium will go into stasis and be viable until it is either destroyed or wakes up.
Stasis storage of mycelium cultures needs no overcomplicating.
THIS, this is the way.
--------------------
 FUCK STIPE 
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rxb
n00b-sabot



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Re: The OFFICIAL Stasis Storage Thread [Re: dna24]
#28291407 - 04/23/23 01:02 PM (9 months, 12 hours ago) |
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i think im going to try using sterile swabs.
they are cheap, they are already sterile, they will soak up an lc.
soak as many as you like in clean(ish) lc, then put them into a vial of DI water. ship anywhere...save 500 an hour if you want to do the work, and potentially legally sell and ship as much as you like (not for me but maybe for you)
i think its easier than paper, and i think its faster to soak up lc than a skewer, i dont think the cotton is enough of a nutrient to be an issue, the only foiling factor might be if a swab didnt get any tissue on it, so run that stir bar at warp5.
this thread is a great inspiration!!! thank you
-------------------- ->$10 FLOW HOOD ALTERNATIVE <- . i cleaned a mold contaminated live culture and saved it. (might have useful applications) [quote]Enlil said: I'd be the guy with thousands of minions doing my bidding and all of the hot women locked in a cage for my use.[/quote]
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SoFlown99
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 1
#28299254 - 04/28/23 07:24 PM (8 months, 26 days ago) |
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Stasis has been achieved 
Thank you for this wonderful writeup and all your help on stasis!
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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: SoFlown99] 1
#28299273 - 04/28/23 07:39 PM (8 months, 26 days ago) |
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Put a few more cultures into stasis today. PC'd 18 of these tubes, cut some long wedges, and dropped em in. Wrap, label, into the rack.

-------------------- LAGM 2023
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SporeJockey
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28337682 - 05/28/23 12:08 AM (7 months, 28 days ago) |
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Just what I needed, this the one. ^^^
Edited by SporeJockey (05/28/23 12:20 AM)
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Way
The


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Re: The OFFICIAL Stasis Storage Thread [Re: SporeJockey] 1
#28337684 - 05/28/23 12:09 AM (7 months, 28 days ago) |
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Quote:
SporeJockey said: .
Excellent first post.
--------------------
That's the way she goes, boys. Sometimes she goes, sometimes she doesn't, cause that's the fuckin way she goes.
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SporeJockey
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Re: The OFFICIAL Stasis Storage Thread [Re: Way] 1
#28337690 - 05/28/23 12:19 AM (7 months, 28 days ago) |
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Lmao, I wanted to save the post but didn’t know how. So at least I can access my threads. 😂
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Screwup
Googles your dumb questions


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Re: The OFFICIAL Stasis Storage Thread [Re: SporeJockey] 1
#28337750 - 05/28/23 02:39 AM (7 months, 27 days ago) |
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Quote:
SporeJockey said: Lmao, I wanted to save the post but didn’t know how. So at least I can access my threads. 😂
Do people have zero idea how to navigate on browsers for bookmarks/favorites? Y’all kill me.
-------------------- 🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 Help US help YOU TEK 2023 Dehydrator TEK
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SporeJockey
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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup]
#28338354 - 05/28/23 02:48 PM (7 months, 27 days ago) |
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Save within the website, not bookmarks…
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Rushing
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Re: The OFFICIAL Stasis Storage Thread [Re: SporeJockey] 1
#28354239 - 06/10/23 08:57 AM (7 months, 14 days ago) |
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Started putting some culture into the stasis library


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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: Rushing] 1
#28355329 - 06/11/23 08:30 AM (7 months, 13 days ago) |
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Love seeing everyone's take on stasis storage! Thats exactly why I created this thread, would love to see more!
-------------------- LITFA LITFA LITFA
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SporeJockey
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Re: The OFFICIAL Stasis Storage Thread [Re: Rushing]
#28355994 - 06/11/23 06:16 PM (7 months, 13 days ago) |
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Could you send a link for those bottles in the last pic? What’s the measurement on them??
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SporeJockey
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye]
#28356004 - 06/11/23 06:18 PM (7 months, 13 days ago) |
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Would it actually go into stasis though since it has more agar to feed on??
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SporeJockey
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Re: The OFFICIAL Stasis Storage Thread [Re: Guerrilla]
#28356034 - 06/11/23 06:30 PM (7 months, 13 days ago) |
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Would that not effectively turn the whole shebang into an LC because of the nutes in the agar?
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Ultravisit0r
apprentice



Registered: 04/03/23
Posts: 55
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Re: The OFFICIAL Stasis Storage Thread [Re: SporeJockey] 1
#28357907 - 06/13/23 03:57 AM (7 months, 11 days ago) |
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Want do you think: Make transfers to water agar plates (no nutes), then transfer to distilled water.
Is that a viable option theoreticaly? Am I missing something?
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DredgeMyEyes
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28357927 - 06/13/23 04:50 AM (7 months, 11 days ago) |
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what's the revival procedure like as far as taking a sample from the vial of distilled water? do you sort of fish it out with tweezers? pin it to side with a scalpel and slide it to the top and remove it? pour out the water completely then remove? I'm probably over thinking it but just would like to know how this part goes lol
Does the water itself become myceliated and can be used to get a plate going?
Best tool and method for each
-Wood sticks -Paper -Wedges
Edited by DredgeMyEyes (06/13/23 04:54 AM)
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Ultravisit0r
apprentice



Registered: 04/03/23
Posts: 55
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Re: The OFFICIAL Stasis Storage Thread [Re: DredgeMyEyes]
#28357954 - 06/13/23 05:32 AM (7 months, 11 days ago) |
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I'd drain the little container and poke little pieces of agar with a scalpel and put it on a new plate. Same thing with paper I guess
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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: DredgeMyEyes] 1
#28358195 - 06/13/23 10:01 AM (7 months, 11 days ago) |
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The medium you use to get myc into your stasis vials makes no difference in the long run. Choose something that you feel comfortable handling.
As far as too much nuts on a medium like an agar wedge, even if the vial does become LC at some point it will run out of nuts and go into stasis. The only concern with this is possibility of it becoming too thick to get out easily but i doubt that would be a concern with just a slice of agar.
-------------------- LITFA LITFA LITFA
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DredgeMyEyes
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28360491 - 06/15/23 06:43 AM (7 months, 9 days ago) |
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What do you guys think about packing a microfuge tube with boiled hydrated wood pieces, wrapped in foil and sterilized in a jar with a sfd and then innocoulated with a few drops of known to be good LC or tiny sample from a plate for long term storage?
Pull out a twig and toss it on a plate to revive
Gonna try it
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Screwup
Googles your dumb questions


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Re: The OFFICIAL Stasis Storage Thread [Re: DredgeMyEyes]
#28360500 - 06/15/23 06:48 AM (7 months, 9 days ago) |
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Why can’t I use filtered or bottled water? Buying a gallon of distilled is so wasteful.
-------------------- 🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 Help US help YOU TEK 2023 Dehydrator TEK
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DredgeMyEyes
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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup]
#28360503 - 06/15/23 06:51 AM (7 months, 9 days ago) |
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I'm fairly certain you can use tap water without issue
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Screwup
Googles your dumb questions


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Re: The OFFICIAL Stasis Storage Thread [Re: DredgeMyEyes]
#28360505 - 06/15/23 06:52 AM (7 months, 9 days ago) |
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Quote:
DredgeMyEyes said: I'm fairly certain you can use tap water without issue
WELL FUCK
-------------------- 🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 Help US help YOU TEK 2023 Dehydrator TEK
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DredgeMyEyes
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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup]
#28360509 - 06/15/23 07:03 AM (7 months, 9 days ago) |
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Probably not 100% ideal just cause there are some dissolved minerals and other stuff but it's fine for agar and LC and all.
Alternatively you could gather some distilled water at home if you look around on the internet there are some methods that involve boiling water in a pot and collecting the condensate in a smaller vessel sitting in the center of the pot I believe.
Someone else will hopefully chime in as I'm no expert lol
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fiddle_head
I'm not the dude, guy



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28360511 - 06/15/23 07:07 AM (7 months, 9 days ago) |
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I’ve been reading on cryovials and centrifuge tubes, think I’m gonna give it a try for my next project. Bout time I got some of these winners in the Rolodex Thanks Donny 🤘
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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup] 1
#28360775 - 06/15/23 12:15 PM (7 months, 9 days ago) |
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I use bottled water just because its what I have on hand.
As for wooden pieces I am finding that putting boiled and sterilized pieces of wood skewers does work but takes a long time for myc to colonize it. I do not know how well it would work just putting a wedge or a bit of LC on it.
-------------------- LITFA LITFA LITFA
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bongoman
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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28366314 - 06/19/23 09:12 PM (7 months, 5 days ago) |
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Let's say I have some spore prints on hand that I don't intend to grow out in the new future.
Would it make sense to start up these spore prints on agar just to get some clean growth to then store using this method?
Would it better to preserve genetics using an agar wedge in water or as a spore print?
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Unikern13
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
#28371733 - 06/23/23 07:26 PM (7 months, 1 day ago) |
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Would plucking invitro pins and dropping them into sterile water tubes work for stasis storage? If no one knows, I’m gonna find out sooner or later because that’s what I ended up doing, but if anyone has any experience that would save me time that would be awesome.
Edited by Unikern13 (06/29/23 11:00 AM)
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fossilshark
DouchebagDonny



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 3
#28405356 - 07/23/23 03:55 PM (6 months, 2 days ago) |
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Threw some Gymnopilus Purperatus and Gandalf in stasis today, decided to switch over to wood sticks as they are way easier to handle then paper.
I wonder how necessary the water even is?

-------------------- LITFA LITFA LITFA
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astra
Mycelia-Of-House-Fungi



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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark] 3
#28410278 - 07/27/23 11:43 AM (5 months, 29 days ago) |
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dH₂O 2ml cryovials

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Leche de
Thick titties


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Re: The OFFICIAL Stasis Storage Thread [Re: DredgeMyEyes]
#28410295 - 07/27/23 11:52 AM (5 months, 29 days ago) |
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Hesitant on the lc part- but it’s a neat idea
-------------------- He need some milk
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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
#28419032 - 08/03/23 04:48 PM (5 months, 22 days ago) |
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Quote:
bongoman said: Let's say I have some spore prints on hand that I don't intend to grow out in the new future.
Would it make sense to start up these spore prints on agar just to get some clean growth to then store using this method?
Would it better to preserve genetics using an agar wedge in water or as a spore print?
That's what I'm doing with all of my spore prints/swabs now. I don't plan on growing for a while, but I'm getting everything germed and put in stasis. Not only is it a great secondary backup, but I think it'll help to speed things up when I'm ready to grow again.
-------------------- LAGM 2023
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side-eye
potate


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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28419036 - 08/03/23 04:50 PM (5 months, 22 days ago) |
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Quote:
fossilshark said: Threw some Gymnopilus Purperatus and Gandalf in stasis today, decided to switch over to wood sticks as they are way easier to handle then paper.
I wonder how necessary the water even is?


How did you like this compared to paper, other than handling? Guess we won't know til revived. Any issue with the sticks floating when you drop em in?
-------------------- LAGM 2023
Edited by side-eye (08/03/23 04:57 PM)
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bongoman
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Re: The OFFICIAL Stasis Storage Thread [Re: side-eye]
#28426261 - 08/08/23 08:15 PM (5 months, 17 days ago) |
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Why use paper etc when the original research study at https://sci-hub.st/10.1139/w08-049 uses 8mm agar punches?
It seems way simpler to just drop a wedge to sterile water no?
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Bigworm



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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman] 2
#28426269 - 08/08/23 08:22 PM (5 months, 17 days ago) |
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Cryo storage works well. I can't say I would trust anything but distilled water with my cultures but I'm sure it's worth a try. I also made mini slants with boiled toothpicks and agar for a shorter storage method compared to the distilled cryo storage.
Edited by Bigworm (08/08/23 08:22 PM)
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KHRIST0PH0R0US
I Am The Walrus


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Re: The OFFICIAL Stasis Storage Thread [Re: fossilshark]
#28440429 - 08/20/23 10:19 AM (5 months, 5 days ago) |
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I was just thinking about this the other day. Advanced methods of storage or even breeding/selecting somehow for hardiness in long term storage. You said "In theory this can lead to changes in the culture that is not desired." - is that covered in the paper you linked to or can you elaborate a little as to what changes or does it just stop at "in theory"? May it be truly random changes that could affect any aspects of it's makeup and manifest itself in an infinite number of ways?
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rizingfire
Mycoticus psychoticus




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Wish I would have known about this back in the day, literally lost gwm iso b cuz the 13yr old print was completely dead but an older version of the original was able to be brought back by Murderlabz so we made iso d from it...tho now we have a nat cross so the new gwmn will be a hybrid. I will save my stuff using cold stasis from now on...way easier
-------------------- aka NHMI
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Screwup
Googles your dumb questions


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Re: The OFFICIAL Stasis Storage Thread [Re: rizingfire]
#28457381 - 09/03/23 04:22 PM (4 months, 22 days ago) |
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Are you just doing marketing for that company lol?
-------------------- 🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 Help US help YOU TEK 2023 Dehydrator TEK
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fahtster
Now With 33%More Faht



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Re: The OFFICIAL Stasis Storage Thread [Re: Screwup] 3
#28486326 - 09/29/23 12:01 AM (3 months, 28 days ago) |
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Getting to be about that time of year for me. Started reviving some dried out grain cultures 8 days ago. I GLC the dry grains and inject them into fresh grains. I can (and sometimes do) go straight to qts but I like to test the cultures first in small jars just to be sure.
Claketi (dried out April 2023)


PE x RW f7 clone (dried out March 2023)


Mak TP (this one is from March 2021)


Fp+ f8 clone (dried out April 2023)


Shit amazes me every time. These new jars will be used to glc to qts for tubs. When I made these new jars, I made a second one of each that will be dried out again for further storage
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Mycomedicine888
Chief Executive Fruitcake



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Loc: in between mushrooms
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Re: The OFFICIAL Stasis Storage Thread [Re: fahtster]
#28498771 - 10/10/23 12:27 PM (3 months, 16 days ago) |
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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?
-------------------- I may have inhaled some of the spores....
    Copelandia cyanescens 2023
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tree frog
eats bugs


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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.
-------------------- 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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DERRAYLD
Constructus


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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) |
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You can just as easily store dried grain vs myceliated grain in a suspension.
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tree frog
eats bugs


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


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

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

Registered: 03/29/19
Posts: 1,494
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
#28509685 - 10/18/23 04:25 PM (3 months, 8 days ago) |
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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 
If someone has better information please share it.
-------------------- LAGM 2.024Stoned Gummys tek (Gummies from sclerotia or mushrooms) *Not just for stones any moreHow to succeed in mycology (and life) - know nothing, read everything, try something, and accept advice. Don't Panic   
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bongoman
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Re: The OFFICIAL Stasis Storage Thread [Re: ReverendMyc]
#28509693 - 10/18/23 04:34 PM (3 months, 8 days ago) |
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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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Nichrome
I'm a torso!


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


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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
#28513424 - 10/21/23 07:41 PM (3 months, 5 days ago) |
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Dry grain spawn FTW.
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SerMantis
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Re: The OFFICIAL Stasis Storage Thread [Re: bongoman]
#28519899 - 10/27/23 08:46 AM (2 months, 30 days ago) |
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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.
-------------------- Wood is good.
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Floret
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Re: The OFFICIAL Stasis Storage Thread [Re: SerMantis]
#28566151 - 12/03/23 12:24 AM (1 month, 25 days ago) |
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These long term experiments are great
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GuyOnaBuffalo
I'm On A Buffalo!

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Re: The OFFICIAL Stasis Storage Thread [Re: SerMantis]
#28566624 - 12/03/23 11:28 AM (1 month, 24 days ago) |
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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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c01h
Aspiring Psychonaut



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Re: The OFFICIAL Stasis Storage Thread [Re: GuyOnaBuffalo]
#28585835 - 12/16/23 06:34 PM (1 month, 11 days ago) |
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3.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) |
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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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c01h
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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) |
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at least 2+ years based of this post
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3.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) |
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Sweet!
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the_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) |
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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? 
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. We'll see these in a decade or so.

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

Thanks to Tree Frog for the link!
-------------------- "Luck favors the observant." - Workman
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SeventhMushroom
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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! 
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
-------------------- LAGM 2024
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MycoSnailCarl
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So at this point do we think LC stored in vials is fine? I’m thinking that it’d be easier/less likely to contaminate if using a vial or LC rather than trying to fish a piece of agar out of water.
Basically, which would be better: putting a small piece of agar in sterile water, or in LC solution?
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Phaethon
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Re: The OFFICIAL Stasis Storage Thread [Re: 3.A.M]
#28614679 - 01/08/24 11:19 PM (19 days, 1 hour ago) |
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Quote:
3.A.M said: 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.
Did you find autoclave pouches that come sterile? Or are you sterilizing the empty pouches first, or just taking a chance since you'll be reviving it on agar anyway and can transfer away from any contaminant that survives dessication?
fahster has a custom Tyvek lid design which I am considering making without any silicone (I would just use them for drying, preferring inoculation without a needle). But if I could dry and store in a single cheap, lightweight, and small container, that would be pretty great.
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3.A.M
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Re: The OFFICIAL Stasis Storage Thread [Re: Phaethon]
#28614720 - 01/09/24 01:18 AM (18 days, 23 hours ago) |
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They’re absolutely not sterile and a couple times I’ve had to transfer away from contam but for ease and practicality they’re amazing, and no I don’t sterilise first but you could easily throw some in a jar and put them through a cycle if you like. I originally got them as an alternative to wrapping home sterilised swabs in foil which isn’t sterile either so grains were just the next natural step, fahts methods are definitely tried, tested and guaranteed successful though if you want to avoid potential contam.
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Phaethon
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Re: The OFFICIAL Stasis Storage Thread [Re: 3.A.M]
#28615920 - 01/10/24 02:06 AM (17 days, 22 hours ago) |
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Cool, thank you! I think I will try that, probably starting by sterilizing the pouches (if that doesn't do something awful to the seal tape).
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3.A.M
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Re: The OFFICIAL Stasis Storage Thread [Re: Phaethon]
#28615940 - 01/10/24 03:08 AM (17 days, 21 hours ago) |
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Literally got some in the pc right now, glad we had this discussion! I don’t use the seal tape anyway, did the first few times but it’s not really designed to come back off so I just masking tape it closed like most people do with swabs. I’ll let you know if sterilisation warps the pouches at all but I don’t imagine it will, I’ve been using autoclaves for years in various industries, those pouches are pretty hardy. You can also get roles of the stuff and heat seal them to any length you like if you want to go that avenue as well👍
Edit: pouches pc’d fine, they were a bit loose in the jar so they sagged down a little and will probably hold that shape but won’t effect function at all (:
Edited by 3.A.M (01/10/24 02:10 PM)
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Phaethon
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Re: The OFFICIAL Stasis Storage Thread [Re: 3.A.M]
#28618006 - 01/11/24 07:26 PM (16 days, 5 hours ago) |
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Nice, thanks for the update!
I've got a couple different pouch sizes in my Amazon cart, along with some bulk desiccant to speed up drying. Will post an update once I've tried it.
Thanks again for the idea! (It's funny - when I first got into the hobby, I was looking for something to enclose things in while pressure cooking, other than foil or rigid containers. I'd seen "autoclave bags" mentioned, and found those almost entirely for the purpose of sterilizing before disposal rather than use. Sterilization pouches and rolls are what I was looking for! I've used polypropylene tubing to enclose stacks of PP petri dishes, but they come out quite wet and stay that way. I'm going to try stacking them in 7.5" x 13" pouches.)
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