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OfflineIGnosticAbhorI
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Storing Strains Long Term
    #3930670 - 03/17/05 03:43 PM (12 years, 1 month ago)

Just bought a package of 100 10ml vac-u-tainers :smirk: :thumbup:

I read on here awhile ago..from somewhere about how you can save your strains long term (years) by using these (airtight)...But I can't remeber if it's spore solution or a LC that you're suppose to put in there....If it's spore solution then i'ma put a whole print per 10ml container (conc. for more syringes)...So that way i can make several syringes from just one container...

So...it's it spore solution or LC?...Since myc. needs certain things to live i think that it'd be just spore solution but would they germinate in just water...(nothing for them to live off so im guessing no)...

Thnkx guys, just wanna double check before I begin storing :thumbup:

-Gnostic


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Offlinescatmanrav
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Re: Storing Strains Long Term [Re: IGnosticAbhorI]
    #3930707 - 03/17/05 03:52 PM (12 years, 1 month ago)

You put mycelium in distilled water, it stops everything. Spores will work too...but if your talking just storing strains really long time, store a dry print, and a mycelium culture in distilled water...


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: scatmanrav]
    #3930712 - 03/17/05 03:53 PM (12 years, 1 month ago)

Alright, my dextrose will be coming shortly so i'll be able to do some LC...Spore prints will be here shortly, Thankx scat :thumbup:

-Gnostic


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InvisibleHolydiver
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Re: Storing Strains Long Term [Re: scatmanrav]
    #3930729 - 03/17/05 03:58 PM (12 years, 1 month ago)

I don't think it necessarily has to be hydrated in order survive long term storage, but it doesn't hurt anything. I revived an old clump of WBS from the bottom of a test tube that had been colonized once upon a time. Growth stopped long ago, and it was completely dehydrated at this point. Not only was it sterile after all that time (placed into test tube as sterile as possible from healthy WBS jar) but growth resumed after being placed in a freshly sterilized jar of WBS.

Dry or hydrated, as long as your sterile technique is there you can store any culture for a long time.

*Edit* The culture was 4-6 months old when restored, not years old.


--------------------
To find a place to live between the negatives and positives.


Edited by Diver (03/17/05 04:22 PM)


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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: Holydiver]
    #3930737 - 03/17/05 04:00 PM (12 years, 1 month ago)

Hmm...Well i'll think i'll test that out and see what happens...But wouldn't myc. tissue die after so long?


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InvisibleHolydiver
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Re: Storing Strains Long Term [Re: IGnosticAbhorI]
    #3930763 - 03/17/05 04:05 PM (12 years, 1 month ago)

Apparently not, it's almost as if the mycelium goes into hibernation, one way or another. I'm guessing there is some survival mechanism at play there.


--------------------
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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: Holydiver]
    #3930787 - 03/17/05 04:10 PM (12 years, 1 month ago)

Hmmm...Well i'll put some Myc. in a vial and i'll post about it in several months or so and see if it works when I give it a little kick (dextrose/water solution)..:smirk:

-Gnostic


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Offlinescatmanrav
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Re: Storing Strains Long Term [Re: IGnosticAbhorI]
    #3930900 - 03/17/05 04:36 PM (12 years, 1 month ago)

"I don't think it necessarily has to be hydrated in order survive long term storage"

4-6 months isnt really to long term..I'm fairly sure to survive decades, it needs to be suspended in distilled water. Im sure myc would survive quite a while without it..but probably not much over a year..thats when I'd start calling it "long term" too. Dont know for sure or not...but I havnt heard of it actually being done yet. I have heard of 10 year old myc stored in distilled water being used though.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Invisibleagar
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Re: Storing Strains Long Term [Re: IGnosticAbhorI] * 1
    #3930928 - 03/17/05 04:44 PM (12 years, 1 month ago)


Vacutainers work great for the storage method below:
~~~~~~~~~~~~~~~~
Sterile water was first used to preserve cultures by Castellani in 1939 (2). Since then, many scientists have used this method; McGinnis et al. in 1974 (3) and Odds in 1991 (4) reported that they were able to maintain viable cultures for more than three years, without degradation.

This technique satisfies many different interests: Castellani's pathogenic fungi, Odds interest was in pathogenic yeast, and McGinnis' interest was in a wide range of fungi, yeast, and bacteria. The distilled water preserved all of them.

Storage Method:
Obtain dram vials from your laboratory supplier and fill them about half full (about 3mL) with distilled water, loosely cap the vials and sterilize them in a pressure cooker for 30 minutes @250 F. Half dram vials or test tubes with screw caps would also work well.

About six milliliters of sterile distilled water is pipetted aseptically into a freshly growing culture. The fragments of hyphae are dislodged by lightly scraping the aerial growth with the same pipette, and the resulting suspension is withdrawn and transferred to a sterile glass vial. Put plenty of inoculum into each vial to insure success. Screw the lid on tight and wrap Parafilm around the top of the vial to make sure it is airtight. When you come back in a few years, you would not want to find that the water had evaporated
.
Store the vials at room temperature away from direct sunlight. A bookshelf or wall cabinet is an excellent place. If conditions deteriorate, and the room should become unbearably hot, the vials can be refrigerated, but that is not normally necessary.

In the distilled water environment, the mushroom culture enters a dormant state, and it is held in stasis
.
The Rude Awakening:
Under aseptic conditions, simply dip a sterile loop into the vial, and streak the mycelia-rich water onto an agar plate. It will start to reanimate on being in a nutrient source and oxygen.

The first four methods keep cultures alive with three items: food, water, and oxygen. If they lack any of these, It's goodbye.

Instead of trying to keep them alive, there is a better way: In sterile distilled water, with no food, oxygen, or minerals. This method was in use almost 60 years ago, but was apparently lost due to lack of communication.

References
(1) M.D.Graham, "A Simple,Practical Method for Long Term Storage of Yeast", Brewing Techniques 5, March/April (1997), pp 58-62
(2) S.Castellani,"Viability of Some Pathogenic Fungi in Distilled Water", Journal of Tropical Medicine and Hygiene 42, pp 225-226 (1939)
(3) M.R.McGinnis,A.A.Padhye,and L. Ajello,"Storage of Stock Cultures of Filamentous Fungi,Yeasts, and Some Aerobic Actinomycetes in Sterile Distilled Water", Applied Microbiology 28, pp 218-222 (1974)
(4) F.C.Odds, "Long Term Laboratory Preservation of Pathogenic Yeast in Water",Journal of Medical and Veterinary Mycology 29, pp 413-415(1991)


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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: agar]
    #3930999 - 03/17/05 05:03 PM (12 years, 1 month ago)

Agar, You're so awesome haha, Even have the references :thumbup:

That's exactly what I was looking for, i'll put this in my fav.s Thankx :grin:

-Gnostic


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InvisibleHolydiver
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Re: Storing Strains Long Term [Re: scatmanrav]
    #3931005 - 03/17/05 05:04 PM (12 years, 1 month ago)

Quote:

scatmanrav said:
4-6 months isnt really to long term..I'm fairly sure to survive decades, it needs to be suspended in distilled water. Im sure myc would survive quite a while without it..but probably not much over a year..thats when I'd start calling it "long term" too. Dont know for sure or not...but I havnt heard of it actually being done yet. I have heard of 10 year old myc stored in distilled water being used though.




I thought about that for a bit too, but thinking further on the topic I don't see a difference between a dried mycelium culture 4-6 months in age, or two years in age. In a sealed environment with no biological action taking place (test tube) no moisture, possibly even no 02, I don't see how the mycelium would/could age or change past 4-6 months.


--------------------
To find a place to live between the negatives and positives.


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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: Holydiver]
    #3931014 - 03/17/05 05:06 PM (12 years, 1 month ago)

Good point Diver...

I'ma still do what I planned on doing, maybe i'll save one longer?

-Gnostic


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InvisibleHolydiver
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Re: Storing Strains Long Term [Re: IGnosticAbhorI]
    #3931081 - 03/17/05 05:20 PM (12 years, 1 month ago)

Yeah you should try it IGnostic, it's something I've been meaning to get back to and test out again.

In short, what I'm trying to say is: What processes, deterioration, or possible aging of the culture could possibly take place past 6 months if they have already occured? Especially in a test tube environment?


--------------------
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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: Holydiver]
    #3931182 - 03/17/05 05:51 PM (12 years, 1 month ago)

:thumbup:

Hmmmm...I've got some myc. growing in jars now..
I'll buy some distilled water soon and get this underway...

Hmmm..Why haven't you tried it yet? No time? Tubes?...I've got 100...soo..:smirk:...Drop me a pm if ur in need :grin:

-Gnostic


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InvisibleMrMaddHatter
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Re: Storing Strains Long Term [Re: agar]
    #3931183 - 03/17/05 05:51 PM (12 years, 1 month ago)

Quote:

agar said:





That there is a nice pic. Makes me drool :tongue:

IGnosticAbhorI, did you order them or buy them locally?

Somebody PM me a place to order these please :crazy:


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Invisibleagar
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Re: Storing Strains Long Term [Re: MrMaddHatter]
    #3931285 - 03/17/05 06:15 PM (12 years, 1 month ago)

Watch EBAY


--------------------


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OfflineIGnosticAbhorI
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Re: Storing Strains Long Term [Re: agar]
    #3931293 - 03/17/05 06:17 PM (12 years, 1 month ago)

Hey hater that was agar's post :smirk:

I can show you in a PM where I got mine but Ebay is typically cheaper :thumbup:
Gl

-Gnostic


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InvisibleMrMaddHatter
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Re: Storing Strains Long Term [Re: agar]
    #3931297 - 03/17/05 06:18 PM (12 years, 1 month ago)

TY! :grin:

though I should've known that :blush:


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Offlinescatmanrav
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Re: Storing Strains Long Term [Re: Holydiver]
    #3931310 - 03/17/05 06:21 PM (12 years, 1 month ago)

Quote:

Diver said:
Quote:

scatmanrav said:
4-6 months isnt really to long term..I'm fairly sure to survive decades, it needs to be suspended in distilled water. Im sure myc would survive quite a while without it..but probably not much over a year..thats when I'd start calling it "long term" too. Dont know for sure or not...but I havnt heard of it actually being done yet. I have heard of 10 year old myc stored in distilled water being used though.




I thought about that for a bit too, but thinking further on the topic I don't see a difference between a dried mycelium culture 4-6 months in age, or two years in age. In a sealed environment with no biological action taking place (test tube) no moisture, possibly even no 02, I don't see how the mycelium would/could age or change past 4-6 months.




Yes, in a sealed environment with no action taking place..I think the issue would be that it still might contain water. And then if you dried it, and placed it in a jar, there would be oxygen in the jar. Overtime that would cause it to break down. I believe the point of the distilled water is to safegaurd it from both of those two points easily. If you were to dry the peice, and store it in an oxygen free environment (co2 with dry ice or something) then it could last for a while.

In fact there is alot of talk in the advanced forum (RogerRabbits got some posts on it, as does Workman) about clones being taken from dried mushrooms that are over a year old. They were rehydrated and started up again, so this would pretty much prove your theory that its possible..the thing is, I think the distilled water makes it easier to store while keeping the most viability possible..


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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InvisibleMrMaddHatter
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Re: Storing Strains Long Term [Re: IGnosticAbhorI]
    #3931319 - 03/17/05 06:22 PM (12 years, 1 month ago)

Quote:

IGnosticAbhorI said:
Hey hater that was agar's post :smirk:




I was trying to talk to both of you in one post :crazy2:

But sure PM me with it :thumbup:


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