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Invisiblejoze
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27387310 - 07/14/21 12:54 PM (2 years, 6 months ago)

Looking forward to this next update :cool:


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: joze]
    #27394525 - 07/19/21 11:06 PM (2 years, 6 months ago)

UPDATE - 15 Days after drop to agar:

Have a look at he OP for more details.

11 Samples in total
5 are showing clear growth
2 are showing probable new growth
4 are not showing any clearly identifiable sign of growth yet.

Conclusion so far: this seems to work better than dropping them straight to LC, "PanCam I" showed growth after 10 days in
LC broth during the previous test but still doesn't show anything here yet. It would be necessary to drop more than just
a couple of kernels onto the plates to get a more average result as some kernels might not be viable or take longer to reactivate.


PanCam "B"

after 5, 9 and 15 days on agar




PanCam "C"

after 5, 9 and 15 days on agar




GymPur

after 5, 9 and 15 days on agar




Papuana

after 15 days on agar




Poplar

bacterial growth showed after 2 days. Mycelium has started to grow as well after 8 days but it's still to thin to get a good shot of it. The visibility
of the plates are poor but you can see some of the hyphae relecting light on the second image. Well its bacterial and not too happy.

images after 8 and 15 days



Edited by Baba Yaga (07/20/21 05:37 AM)


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27423246 - 08/10/21 07:50 PM (2 years, 5 months ago)

Updated OP with the following:

Revival Test Part 4

This will be the last test trying to revive dehydrated grain spawn. This time I will try to revive the grain that hasn't shown any grows in
the previous two tests. What I'm doing different this time is putting 10+ grains on each dish to get a better chance of having viable kernels
on the plates. I still got some more dried cultures that I haven't tested yet but will leave that for sometime later. I might repeat this from
scratch and try a more controlled drying method in a chamber with a salt that will buffer the humidity in the chamber to a certain level so all
grains will dry under the same conditions....maybe but I am keen on trying some other things as well and there is only so much time so we'll see.


Grains dropped to agar on August 11th: 

Formula: 0
*Added a second Sub culture, LME and BRF indicating the LC nutrient use for the culture.

You find the updated table in the first post.


Edited by Baba Yaga (09/01/21 12:37 AM)


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27432351 - 08/17/21 08:56 PM (2 years, 5 months ago)

Update Revival Test #4: 7 days after drop to agar

3 out of ~50 kernels of the Subaeruginosa spawn made with BRF LC are showing new growth after 7 days on agar.



Edited by Baba Yaga (08/28/21 05:23 PM)


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27446673 - 08/28/21 05:21 PM (2 years, 4 months ago)

Update Revival Test #4: 18 days after drop to agar

Sub BRF

3 out of ~50 kernels of the Subaeruginosa spawn made with BRF LC are showing new growth after 7 days on agar. Second image of fully colonized plate after 18 days.




Cube

Finally after 18 days one of the cube kernels is showing growth. This spawn was dehydrated 1 year and 7 month ago.



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OfflineSingularFusion
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27451606 - 08/31/21 10:32 PM (2 years, 4 months ago)

interesting Baba

:takingnotes:

:rockon:


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InvisibleGuerrilla
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: SingularFusion]
    #27452052 - 09/01/21 10:17 AM (2 years, 4 months ago)

Great stuff, commenting so I can come back to this!


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InvisibleBlack and gifted
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Guerrilla]
    #27455778 - 09/04/21 10:00 AM (2 years, 4 months ago)

Same, cool stuff.

I take long breaks from the hobby, and I'm interested better storage. I recently attempted to start back up and realized the culture slants long forgotten in the back of my cabinet were dried up. I had no luck reviving.


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Black and gifted]
    #27493912 - 10/05/21 06:19 PM (2 years, 3 months ago)

Last up date:

The cube spawn that was in dry storage for 1 year 7 month pinned.

I'm very glad this clone came back to life as I lost my other cultures of it.



For final conclusion see OP.

TL;DR: wished it worked better but that is probably due to process flaws, a more refined dehydration method of the grain spawn will
probably yield better results.

Moving on to other long term storage methods now......when I find the time....

:sun:


Edited by Baba Yaga (10/07/21 12:36 AM)


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InvisibleIchigo Kurosaki
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27503703 - 10/13/21 10:21 PM (2 years, 3 months ago)

So let me clarify, those are completely dry grains? Or dry to the point of forcing the myc into a hibernation like state? Which seems more likely.


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Ichigo Kurosaki]
    #27503759 - 10/14/21 12:15 AM (2 years, 3 months ago)

Some of the grains I took out after 2-3 weeks and some I forgot about and left them in there for a couple of month.
I actually have no idea how dry the individual batches of grain were that, my fridge can dry small mushrooms fairly
quickly to the point where they are cracker dry. So I guess some where totally dry and some might have had still some
moisture in them.

If I would do it again I would, as said before, do it in a chamber with controlled humidity at ~10% or even less, but I don't
have the time to figure that one out, sounds more like a retirement project :wink:


In theory the mycelium should survive even when completely dried out as some of the papers suggest that I read, its just a matter
of doing it the right way. In one article I read that they had the myc grow over sterilized paper tissue that was placed on a
petri dish and dried the paper for long term storage, that I might try with a couple of species, already sterilized some paper.


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InvisibleIchigo Kurosaki
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27504375 - 10/14/21 04:09 PM (2 years, 3 months ago)

Interesting, this makes me ponder if someone could biopsy Enigma or clones for example. That's dried completely. As this could make it much more convenient to obtain. But The biggest difference is most dehydrators are far beyond 109F. Which is when spores\myc start to die. At least so I've read. You know how that goes on here, with a grain of salt lol. So for said experiment to even be done. Someone with a culture and a means to dehydrate at low temps would be needed. A fridge I suppose. It would be pretty epic if possible. But from what I've seen, people using traditionally dried fruit have issues with contams. It would definitely be a challenge but hey never know might be the next breakthrough right.

Good luck to anyone who attempts this, I'll be crossing my fingers :mushroom2:


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Ichigo Kurosaki]
    #27504520 - 10/14/21 06:10 PM (2 years, 3 months ago)

Yeah the challenge is to dry the samples while keeping them sterile as the mycelium will take a while to
come back to life. Freeze drying might be a good way to do if you have such a machine at hand but
you still will have to have the samples inside of a filtered container/jar, no idea how well this might work.

Gotta keep it sterile and no heat.....wished I had more time.


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Offlineyas champ
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: gt40]
    #27538859 - 11/11/21 07:03 AM (2 years, 2 months ago)

Hello baba yaga,
is it possible to know the composition of LC please? is it water and honey only? or is there something else?
thank you


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: yas champ]
    #27538998 - 11/11/21 09:15 AM (2 years, 2 months ago)

Light malt extract is all I use for LC, concentration for these tests was 1%.


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Offlineyas champ
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27542526 - 11/14/21 02:06 AM (2 years, 2 months ago)

what do you mean by 1%? 10 g/l malt extract?
do you use distilled water or will tap water do?
thanks :smile:


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: yas champ]
    #27542533 - 11/14/21 02:32 AM (2 years, 2 months ago)

Yes, I used 10g/l for the tests, TBH after I did these tests I thought that I could have dropped the nutrient concentration down to what I usually use for LC which is 0.1% so 1g/l.

Using distilled or at least bottled water cause it makes a clearer broth than my tap.


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Offlineyas champ
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27542675 - 11/14/21 06:38 AM (2 years, 2 months ago)

Thank you for your answers :le sourire:,
one last question :sourire:: does the quality of the mushroom play a role in the culture of the mycelium?  make a culture on agar plate or LC with a mushroom resulting from a 1st generation and another of a n generation will have the same result? (growth speed, color...) ?


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InvisibleBaba Yaga
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: yas champ]
    #27542925 - 11/14/21 11:10 AM (2 years, 2 months ago)

Sorry, I don't think that I really understand your question? That's probably because I don't know where you are coming from and where you
are planning to go with this.

It would help to explain what the bigger picture is, like how are  making your cultures and how are you planning to use the dehydrated cultures.

You read that the experiments gave me mixed results and about half the cultures wouldn't come back to life?


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OfflineSingularFusion
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Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga] * 1
    #27543593 - 11/14/21 08:52 PM (2 years, 2 months ago)

I also had trouble reviving things

Was dehydrated tiny (3 x 3mm approx) chips of wood from some woodlover stuff I'm doing

Dehydrated in a filtered little mycobox like what the kits come in, I left it about five months at ambient conditions to dry out and then put to malt yeast agar plates for revival. I poured those a bit hot so there would be some added condensation to wet the chip once it was transferred in

Azure revived but cyan would not, even though you can see mycelial white on the chips, it just won't jump off. Been about six weeks and plates are otherwise spotless. I am keeping them around to see if anything happens but it looks like a fail on revival for me also... Well 50% success rate anyway

I think the method itself has some promise and potential, but there must be a knack to the dehydration that allows revival, something I'm not fully understanding, to make it work right :shrug:

I look forward to seeing what you can figure out in the future Baba


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