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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Adas] * 1
    #28181029 -

It is easier to get scrab some cultures off of a wedge than off of a fruit I could imagine :shrug:


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I am not crazy, I prefer the term mentally hillarious.

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Shrimps] * 3
    #28181035 -

also i feel like a higher risk for contam. probably would be fine with inner tissue but the wedge would give me more security personally


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: hazyhorse] * 1
    #28191262 -

Did anyone notice that the wedges grew out some myc around them after transfering into the vials? It's faint and diffuse growth but I can see it. I mean I kinda expected it, just wanted to ask others if it's normal, just to be sure.


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Shrooms are always bluer on the other side...

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Adas] * 3
    #28191298 -

I am prepping my first vials on Sunday. Want to save up my pearly gates, starry night and Lions Mane for long term^^ I can tell you then when they've sat in there for a while ;-)


--------------------
I am not crazy, I prefer the term mentally hillarious.

Agar - The way to go!
Clean spawn checklist
Proper Surface Moisture
Recognizing and dealing with contamination
πŸ…ƒ πŸ„΄ πŸ„° πŸ„Ό    πŸ„² πŸ„» πŸ„Έ πŸ„½ πŸ„Ά πŸ…† πŸ… πŸ„° πŸ„Ώ

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Adas] * 3
    #28191299 -

That's what all mine did. Ate the sugar in the transfered wedge I figured.


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Links For Beginners And Beyond:mspoil:

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Adas] * 3
    #28191300 -

Yeah I saw that in mine adas! Very faint but prolly just using up the remainder of the agar nutrient..

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: kirkeng] * 3
    #28191322 -

will also add mine have done the same thing. they look like little ghost blobs. didn’t seem to matter much when reviving the culture!


--------------------
you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================

πŸ…ƒ πŸ„΄ πŸ„° πŸ„Ό    πŸ„² πŸ„» πŸ„Έ πŸ„½ πŸ„Ά πŸ…† πŸ… πŸ„° πŸ„Ώ

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: hazyhorse] * 3
    #28191389 -

Awesome! Thanks gang :hug:


--------------------
Shrooms are always bluer on the other side...

Work hard, play harder, trip the hardest.

Simple Cloning TEK by Adas
Semi-Sterile TEK (SS-TEK)
Growing Panaeolus bisporus - The Low-Tech approach
Exotic Mycelia Surviving Winter - Creating IceBreaker Variants!
My First Panaeolus Cyanescens Grow!
Psilocybe azurescens photo compilation 2020
Azurescens in June!

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Adas] * 2
    #28191554 -

Hello shroomerites,
i do something quite similar.
I use hplc vials, they are autoclavable and have a septum in the lid, so one can poke through with a syringe needle.
i autoclave them empty.
instead of wedges i use LC, in which i exchanged the Nutrient Media with dH2O.
its a bit more laborious, but one can have a lot of concentrated material in a tiny 1,5ml vial, and just suck it up with a syringe later on by poking the lid.

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Re: 'Cryovial Tek': Long-Term Storage of Cultures with Cryogenic Vials [Re: Mycoplex] * 1
    #28342664 -

:threadmonitor:


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:ancientaliens:

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Re: 'Cryovial Tek': Long-Term Storage of Cultures with Cryogenic Vials [Re: astra] * 1
    #28452496 -

So, is using distilled water an absolute must? Is demineralized/deionized water not good enough? Very difficult to find distilled around here, but demineralized is literally everywhere


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Re: 'Cryovial Tek': Long-Term Storage of Cultures with Cryogenic Vials [Re: odissey] * 3
    #28452502 -

Demineralized and deionized water both leave behind trace biological and particulate matter that distilled removes. Since we are going to sterilize the water any way, you might think that this is good enough. However, although the sterilization process will kill any nasties, it will not remove them or their byproducts. So you will not end up with true stasis until any live mycelium consumes or ignores any remnant materials.

tldr: Ideally, no. For short term use, probably. For long term storage, maybe.

p.p.s. They sell water distillers for home use for $50 and up. Not sure how practical that is, though. Amazon has them.


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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 (08/30/23 05:34 PM)

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Re: 'Cryovial Tek': Long-Term Storage of Cultures with Cryogenic Vials [Re: ReverendMyc] * 1
    #28452548 -

ReverendMyc said:
Demineralized and deionized water both leave behind trace biological and particulate matter that distilled removes. Since we are going to sterilize the water any way, you might think that this is good enough. However, although the sterilization process will kill any nasties, it will not remove them or their byproducts. So you will not end up with true stasis until any live mycelium consumes or ignores any remnant materials.

tldr: Ideally, no. For short term use, probably. For long term storage, maybe.

p.p.s. They sell water distillers for home use for $50 and up. Not sure how practical that is, though. Amazon has them.


Thanks for the reply! Very clear :thumbup:


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mandrake Erpelmann] * 1
    #28543677 -

Mandrake Erpelmann said:
Hello shroomerites,
i do something quite similar.
I use hplc vials, they are autoclavable and have a septum in the lid, so one can poke through with a syringe needle.
i autoclave them empty.
instead of wedges i use LC, in which i exchanged the Nutrient Media with dH2O.
its a bit more laborious, but one can have a lot of concentrated material in a tiny 1,5ml vial, and just suck it up with a syringe later on by poking the lid.


My method does not seem to work so well...tried reviving Tidalwave 2 culture that i prepared  as described a year ago, and could not bring it back to life.


Any ideas why?

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mandrake Erpelmann]
    #28655549 -

Mandrake Erpelmann said:
Mandrake Erpelmann said:
Hello shroomerites,
i do something quite similar.
I use hplc vials, they are autoclavable and have a septum in the lid, so one can poke through with a syringe needle.
i autoclave them empty.
instead of wedges i use LC, in which i exchanged the Nutrient Media with dH2O.
its a bit more laborious, but one can have a lot of concentrated material in a tiny 1,5ml vial, and just suck it up with a syringe later on by poking the lid.


My method does not seem to work so well...tried reviving Tidalwave 2 culture that i prepared  as described a year ago, and could not bring it back to life.


Any ideas why?


Have you tried doing the same experiment, with the same culture, but transferring via agar piece instead of the LC-based method you describe?  If you do them side by side and see that one works better that might narrow in on it.

I'd expect the LC to behave the same as the wedge, so maybe this was a luck thing and it did not work out with this one vial.  Did you take backups on multiple vials, and did they all behave this same way too (none revived)?


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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Mycoplex] * 1
    #28705353 -

I lost a bunch of cultures on agar slants and plates due to my fridge freezing them. Nothing really valuable, but my first reasonably good cube clones and early backups of some gourmet cultures. Mainly I was stupid and didn't have them in an insulated container, but this thread inspired me to stop bothering with the fridge entirely. Cryovials are so cheap, the freezer boxes are equally cheap and make it super easy to find what you want without rummaging around (assuming you keep good notes - of course good labels are also necessary), and it's really easy to prepare a ton of these and drop wedges in whenever it seems worthwhile. Thanks!

I found I could easily fit 13 of the 2 mL tubes upright in a 4 oz mason jar. Once sterilized (I used an unmodified PP lid on the jar, slightly loose), it was also pretty easy to remove one at a time to tighten each vial's lid without knocking any over. (Could even leave them as is, in the jars, but they take up a lot less space in a freezer box.) Pretty easy to process a ton at a time this way, without using any foil or paper towels.

I'm tempted to try a run with them in a freezer box, which is also polypropylene. Just a little worried about pooling water in the box, though I guess if that happened, you could just tip it over with the box lid unclasped after tightening all of the vial lids.

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: Phaethon] * 2
    #28734153 -

Are you guys scraping your micro wedges off your scalpel with the edge of the container? I’m getting ready to do my first batch of backups into cryovials soon so I’m starting to think about my procedure.

Was thinking about doing some Josex pokes into the vials but then I’m thinking it may be hard to recover once I’m finally going to recover them

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: TPython] * 3
    #28734186 -

My wedges are not so micro, but yes, I do kid of use the inside of the cryo-tube to get the piece off of the scalpel if needed. Also kind of use the side to press against when retrieving a piece when restarting a sample.


--------------------
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] * 1
    #28734244 -

Oh damn, those are pretty big. I was reading something saying that you should try and keep the volume of water 50x bigger than the sample so that it goes into dormancy faster but maybe that isn’t very important in practice.

Have you had good luck reviving wedges that large?

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Re: Tek: Long-Term Storage of Cultures with Cryogenic Vials (Cryovials) [Re: TPython] * 3
    #28734311 -

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.


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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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