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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: ReverendMyc] * 1
    #28734338 -

Would a low nutrition solution like 0.1% help spread the mycelium through the
water or does the agar wedge already do that over time?


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Using Ziplocs (Ziplocks) as Fruiting Chambers

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Psilosion] * 2
    #28734363 -

ReverendMyc said:
So far I have had 100% success, but I have only done 10 or so and none more than a 18 months old yet. Now I actually do more with a whole plate chopped up into a small media bottle half full of sterilized distilled water. Same concept on a larger scale with the added benefit of pouring a little directly to grain as an li. I keep one as master and one as usable LI. Any time I run low on LI, I make a plate from the master and when it grows out clean, chop it up and into another sterilized water media bottle.


Very cool, thanks ReverendšŸ‘

I like the plate and media bottle idea. Originally I was planing on backing up a few wedges to 50ml centrifuge tubes but space wise the cryovials make so much more sense. Figured I could always do multiple vials of the same culture and still take up less of a footprint than those fat 50mls

I just donā€™t get how Mycoplex cuts those perfect lil agar cubes, dudes a damn surgeonšŸ˜„

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Psilosion] * 2
    #28734484 -

Psilosion said:
Would a low nutrition solution like 0.1% help spread the mycelium through the
water or does the agar wedge already do that over time?


For this particular tek the idea is to suspend growth, so adding nutrition would be antithetical. It for sure works as an lc, but here we are trying to put genetics into stasis to preserve them. Any added nutrition risks undesirable foreign growth to contaminate our precious cultures.

The LI I mentioned does supply the remaining nutrition in the agar wedges and the myc does continue to grow and break off for a while. But the wedges don't really dissolve and release nutrients. Hopefully the mycelium eats the remaining nutes and then goes dormant until we give it something new to chew on.

TPython said:
I like the plate and media bottle idea. Originally I was planing on backing up a few wedges to 50ml centrifuge tubes but space wise the cryovials make so much more sense. Figured I could always do multiple vials of the same culture and still take up less of a footprint than those fat 50mls

I just donā€™t get how Mycoplex cuts those perfect lil agar cubes, dudes a damn surgeonšŸ˜„


Right? I can't even see the tiny bits he is able to snatch.
Ya, you can't beat the tiny cryovials for optimizing storage space. I still use them for that and they are great to mail in a bubble envelope as well. They are all kinds of handy.

Reminds me of a package I need to get out now that the weather has started to settle down in the northern hemisphere.


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Stoned Gummys | BRF Pucks | Primo Reusable Plates | Easy LI 4 Preserv & Propo
"Psychedelics are powerful substances. Nothing that powerful is completely safe... and nothing completely safe is that powerful!" - Abigail Calder at ALPS 2023
Don't Panic

Edited by ReverendMyc (04/11/24 11:47 PM)

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: ReverendMyc] * 3
    #28735427 -

UTB has awaken from a year in a cryo.

Thanks psi!



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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Atomsplit] * 3
    #28735833 -

I'm scared.. of whoever is in that plate's reflection.. :scaryshroom:


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Adas] * 2
    #28736238 -

miles morales :biggrin:
The two spider verse movies were really good. If you haven't seen em I recommend you do.


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Atomsplit] * 2
    #28754483 -

Made my first 4 of these, just backing up some clean agar dishes before I put them to grain.


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Using Ziplocs (Ziplocks) as Fruiting Chambers

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 4
    #28778587 -



Going to give these a try for long term storage and of course sharing.

:awesomenod:


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Smellyhobbit said:
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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: HILLBILLY OUTLAW]
    #28781790 -

Lookin good fellas!


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28794382 -

How many agar wedges are you transferring to a single vial?

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: bongoman] * 1
    #28794438 -

I do one elongated rectangle a good portion of the size of the vial.

I would like to hear what others do as well.



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Stoned Gummys | BRF Pucks | Primo Reusable Plates | Easy LI 4 Preserv & Propo
"Psychedelics are powerful substances. Nothing that powerful is completely safe... and nothing completely safe is that powerful!" - Abigail Calder at ALPS 2023
Don't Panic

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: ReverendMyc] * 2
    #28794542 -

I usually put 2-4 little squares in:


It doesn't take much to make a new plate, and an inoculation loop is a perfect tool for fishing out a square of agar. Using a vial multiple times probably increases the risk of contamination, but I think the risk is low (especially with sterile flow) and I always make backup vials.


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Phaethon] * 2
    #28794864 -

I only put one little triangle in mine cause I thought the point was to limit the amount of nutrients in the vial so the mycelium goes into a more stasis mode for storage. I would think the more nutrients you put in there then the more the mycelium would grow.


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: ReverendMyc] * 1
    #28794916 -

ReverendMyc said:
I do one elongated rectangle a good portion of the size of the vial.

I would like to hear what others do as well.




I got a few cultures in the mail in these tubes recently and immediately ordered a case.
I have no idea why I didn't do this years ago. Were we doing this years ago???


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Cultivating Panaeolus cyanescens


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Asura] * 2
    #28794993 -

lipa first mentions it about 12 years ago, but I only started 2 years ago after reading this OP.


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Stoned Gummys | BRF Pucks | Primo Reusable Plates | Easy LI 4 Preserv & Propo
"Psychedelics are powerful substances. Nothing that powerful is completely safe... and nothing completely safe is that powerful!" - Abigail Calder at ALPS 2023
Don't Panic

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: ReverendMyc] * 3
    #28795009 -

I use to use the little test tubes with injector ports to keep
LC samples, but somehow I missed this one. Slants...never again.


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Cultivating Panaeolus cyanescens


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Asura] * 1
    #28832446 -

Awesome thread. Reading through this, I didnā€™t see a lot of people mentioning how long they have stored & revived cultures.

I saw the link to the study that claimed up to 20 years. But whatā€™s your guys longest timeframe with success??

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: StrappedTurtle] * 1
    #28837049 -

StrappedTurtle said:
Awesome thread. Reading through this, I didnā€™t see a lot of people mentioning how long they have stored & revived cultures.

I saw the link to the study that claimed up to 20 years. But whatā€™s your guys longest timeframe with success??


I've personally revived cultures with this method that are over a year old (they stayed in the vial for a year and got revived on agar), and currently I have multiple vials that are now getting closer to 2 years old and am confident they would revive as well.  I am not doing cult. at the moment but the next time I'm planning on reviving a culture of "Tasmanian" cubensis that I saved using this method a couple years back.

I'm sure there is an upper limit on how long they do indeed last, but for practical purposes I think it's long enough to meet whatever use case you have in mind for 1, 5, 10 or more years down the line.

Also remember that you can take multiple backups of the same culture, so by saving multiple vials you can be a little more assured that even if one or more kicks the bucket down the line for some reason, you'll have other vials available.


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 2
    #28907717 -

Hey guys, as of earlier this week I've also successfully revived a cubensis culture that I stored using this method back in February 2022 which was a few months before I authored this thread.

The culture revived on agar successfully after about 4-5 days which admittedly is longer than it would take to see growth if you were to inoculate agar fresh LC, but it's not that much slower considering what's being done.

I also used a small operational 'trick' during this resurrection which I wanted to share.  It removes the need for a scalpel, inoculating loop, syringe, or other tools when restoring a culture saved with this tek.

Before inoculating the agar, I vigorously shook the cryovial that contained the sample from 2022.  I disinfected the outside of the vial during this process.  When shaking the vial this way, it will invariably break up microscopic (or larger) bits of the agar that haven't already done so on their own over time. 

After this, all I did was gently pour a small amount of the liquid in the vial into each agar cup without using any tools, just gently and quickly leaning the vial over an agar cup.  This is tricky because it's easy to accidentally pour out the whole contents but even if you do so you should still end up with one agar dish/cup that stands a good chance of restoring.  Otherwise, it should just take a couple drops in each agar dish.


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28908772 -

Mycoplex said:
...I also used a small operational 'trick' during this resurrection which I wanted to share.  It removes the need for a scalpel, inoculating loop, syringe, or other tools when restoring a culture saved with this tek.

Before inoculating the agar, I vigorously shook the cryovial ...  When shaking the vial this way, it will invariably break up microscopic (or larger) bits of the agar that haven't already done so on their own over time. 

After this, all I did was gently pour a small amount of the liquid in the vial into each agar cup without using any tools...


Excellent result.

I found the same thing on a larger scale with making easy LI inspired by this thread. Nearly the exact same process, but using media bottles instead of cryo vials and nearly whole agar plates. A good shake before pouring and then I can go straight to grain. My jars usually finish in 7 to 12 days.

I still do cryo vial for convenient long term storage and keeping an off-site backup for disaster recovery.


--------------------
Stoned Gummys | BRF Pucks | Primo Reusable Plates | Easy LI 4 Preserv & Propo
"Psychedelics are powerful substances. Nothing that powerful is completely safe... and nothing completely safe is that powerful!" - Abigail Calder at ALPS 2023
Don't Panic

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