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Invisibleflyhighfunguy


Registered: 09/13/17
Posts: 1,553
Do you think its still viable? (8.5 month old refrigerated LC)
    #25579017 - 10/30/18 03:22 PM (6 years, 2 months ago)

Hi everyone,

Ive got about 1cc of liquid culture left in a syringe from about 8.5 months ago. Its been in a refrigerator for at least 7 months of that time.

Do you guys think i have a good chance of getting some growth with this syringe? Im trying to decide if its worth trying out or not.

Thanks


--------------------
If you need any help achieving proper field capacity, please check out this video. I hope it helps you guys out!

https://www.youtube.com/watch?v=HskufFlXEo4

https://www.shroomery.org/forums/showflat.php/Number/25087128/page/1

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OfflineWeavieWonder
Time Travel Sucks
Male User Gallery


Registered: 08/18/16
Posts: 1,455
Loc: USA Flag
Last seen: 1 year, 22 days
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: flyhighfunguy] * 1
    #25579088 - 10/30/18 03:50 PM (6 years, 2 months ago)

This is a tampanensis culture that I just tested on agar.  The LC had been stored in the fridge for eight months.


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Offlineverum subsequentis
seeker of truth
I'm a teapot User Gallery


Registered: 03/22/16
Posts: 8,732
Last seen: 2 years, 7 months
Trusted Cultivator
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: WeavieWonder]
    #25579103 - 10/30/18 03:58 PM (6 years, 2 months ago)

i've been wanting to do some tests to see how long an lc would last in cold storage but haven't gotten around to it yet. i'll be curious to see what you find

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OfflineDreamReality
MycoPilot
I'm a teapot


Registered: 03/31/09
Posts: 477
Last seen: 4 months, 7 days
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: verum subsequentis]
    #25579206 - 10/30/18 04:53 PM (6 years, 2 months ago)

Seems worth


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Dream's Mono Flip Tek


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Offlineteladi
FUNKSOULBROTHER


Registered: 06/27/17
Posts: 1,189
Loc: South Africa Flag
Last seen: 1 year, 5 months
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: verum subsequentis]
    #25580563 - 10/31/18 02:38 AM (6 years, 2 months ago)

I'm sourcing some tubes* to do exactly this. I'm seeing a lot of different methods for LC storage in labs. Most seem to freeze them with varying levels (5%-50%) of sterile glycerol, sometimes with a paraffin/kerosene/mineral oil layer on top to slow desiccation or stop biological activity. 

*Still trying to figure out which tubes and find a protocol. Culture, test, micro-centrifuge. I'm leaning towards the micro-centrifuge tubes. You don't really need to store much, and at ~$10/500 for 2ml they aren't pricey.

A protocol that I've considered, but requires investment into some lab equipment, specifically a pipettor and possibly a centrifuge:

1. PC 2ml micro-centrifuge tubes.
2. Add in 1.0-1.5 ml broth per tube.
3. Inoculate Josex style.
4. Grow for however long it takes. The lack of GE may be a problem if they are capped, in which case they'd need to be grown in a container with filtered air.
5. Add glycerol. Mix.
6. Freeze the tubes, maybe with some kerosene/paraffin or mineral oil.
7. Thaw the tubes slowly.
8. Centrifuge and collect the deposited culture, or pipette out the culture and broth.
9. Inoculate a large prepared broth with the culture.

In a container with the space of less than a sandwich, you can store 100 difference cultures.

Pie in the sky type of thing, though. I'll probably run my tests with test tubes sealed with rubber bungs.

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Invisiblenube424
Male

Registered: 12/03/17
Posts: 6,063
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: teladi]
    #25580603 - 10/31/18 03:34 AM (6 years, 2 months ago)

That's pretty interesting...

And how would an lc do that's been stored at room temp for 3 months?

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InvisibleTimehole
 User Gallery


Registered: 04/28/12
Posts: 156
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: teladi]
    #25580609 - 10/31/18 03:39 AM (6 years, 2 months ago)

Jurassic park comes to mind with talk of those micro-centrifuge tubes lol
I think the freezing temps would likely kill any cube though..

@OP I had a 3 year old Reishi LC in my fridge that recently surprised me when i put it on grain and it was still alive. I was about to throw it out but figured might as well give it a go. Even my refrigerated agar dishes of the same culture had failed after that time, I think mainly because they had dried out quite a bit. Definitely worth a try!

In this thread eatyualive mentions and uses an 11 year old Oyster LC that is still rocking. :rockon:
https://www.shroomery.org/forums/showflat.php/Number/21744280/fpart/all/vc/1

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Offlineteladi
FUNKSOULBROTHER


Registered: 06/27/17
Posts: 1,189
Loc: South Africa Flag
Last seen: 1 year, 5 months
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: Timehole]
    #25580613 - 10/31/18 03:45 AM (6 years, 2 months ago)

Should be just fine. There are a few pics of forgotten LCs growing a mat at the top surface and pinning.


Quote:

Timehole said:
Jurassic park comes to mind with talk of those micro-centrifuge tubes lol
I think the freezing temps would likely kill any cube though..





Plenty of people here freeze their slants. Fungi are awesome!

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Invisiblenube424
Male

Registered: 12/03/17
Posts: 6,063
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: teladi]
    #25580616 - 10/31/18 03:48 AM (6 years, 2 months ago)

The one I have in my closet has a Mycelium mat on top. I'll give it a shot this weekend

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OfflineRalphieRamone
Illustrious Nemophilist
I'm a teapot User Gallery


Registered: 08/12/18
Posts: 104
Loc: Jupiter
Last seen: 4 years, 4 months
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: verum subsequentis]
    #25580991 - 10/31/18 08:24 AM (6 years, 2 months ago)

Quote:

verum subsequentis said:
i've been wanting to do some tests to see how long an lc would last in cold storage but haven't gotten around to it yet. i'll be curious to see what you find



I had a Golden Teacher LC made with honey that I had kept in the fridge for 2 years. Never let my girlfriend throw it away haha.
This year I used it on 6 cakes and only 2 contam. The rest produced moderately..think I got like 7g total from them.
If you've kept it from contaminating and at a constant low temp it should still be viable IME!
I've also just used a 3 year old B+ print to make syringes and they worked great!

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Invisibleflyhighfunguy


Registered: 09/13/17
Posts: 1,553
Re: Do you think its still viable? (8.5 month old refrigerated LC) [Re: RalphieRamone]
    #25621144 - 11/17/18 02:48 PM (6 years, 1 month ago)

Whoops, i accidentaly forgot about this thread after i made it lol.

I appreciate all the responses tho. It seems like the majority of you think it should still be viable :rockon::awedance:  I'll most likely be putting it to agar within the next few weeks or so, so ill be sure to keep this thread updated with what happens.

Thanks for the help guys :super:


--------------------
If you need any help achieving proper field capacity, please check out this video. I hope it helps you guys out!

https://www.youtube.com/watch?v=HskufFlXEo4

https://www.shroomery.org/forums/showflat.php/Number/25087128/page/1

Extras: Filter Print Post Top
Jump to top Pages: 1

Shop: Myyco.com Golden Teacher Liquid Culture For Sale   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Kratom Powder for Sale   North Spore Cultivation Supplies   Bridgetown Botanicals CBD Concentrates   MagicBag.co All-In-One Bags That Don't Suck   Mushroom-Hut Liquid Cultures


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