Home | Community | Message Board

World Seed Supply
This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: North Spore Bulk Substrate   Mushroom-Hut Liquid Cultures   Left Coast Kratom Buy Kratom Extract   Kraken Kratom Red Vein Kratom   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Kratom Powder for Sale   Bridgetown Botanicals CBD Concentrates   Unfolding Nature Unfolding Nature: Being in the Implicate Order

Jump to first unread post Pages: 1 | 2 | 3  [ show all ]
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Long Term Culture Storage On Dehydrated Grain + Revival Tests * 7
    #26943501 - 09/19/20 04:01 AM (3 years, 4 months ago)

Long Term Culture Storage on dehydrated Grain

This is an Experiment
about storing cultures long term (5-10 years). The storage method is dehydrated grain spawn and a first revival attempted after
8 month of dry storage turned out successful which lead me to do this write up. Further tests to revive the same cultures will be carried out in
intervals of a year from now on. This is nothing new, I just want to show a practical/easy way to produce dehydrated grain spawn on a small scale.
It's still early days for my experiment, but the proof of concept is there and cultivation performance as well as long term performance of
the cultures after a few years in storage will have to be determined which is especially of importance for clones and isolates but as long as you are
getting a few fruits to print you are up and running again and maybe reviving 5 year old grains turns out to be easier than getting 5 year
old spores going.

TL;DR: There is a final conclusion on this experiment at the end of the post for those who want to know how this turned out first but feel free
to read through the whole post if you are inclined to try it yourself so you can do things differently and go beyond the point where I ended.



It was brought to my attention by Pastywhyte that drying the grains in a fridge will not work as well or not at all in tropical climates (hot & humid)
which I wasn't aware of. The model and age of your fridge will also have an impact on the efficacy of this method I guess. In these cases
you might want to look into storing cultures in distilled water as this seems to be a good option as well, even at room temperature.
Long explanation cut short: This method worked for me so far and I am not claiming it will be for everyone but feel free to experiment :cookiemonster::elmo:



Well I was curious about alternative methods to store cultures long term. Currently I do use just petri dishes and liquid sample containers which I use as "slants".
This is all good and works pretty well although I would like to reduce my workload eventually by not having to re-slant all my cultures every year or two.

Through some research I found the following two methods which seemed to be the easiest to achieve with what I already have and do:

- storing mycelium in sterilized water or a low nutrient broth

- storing mycelium as dehydrated grain spawn

I mean there were more options mentioned such as storing in sterilized sand and perlite (both soaked in a low nutrient broth), frozen, grown
out on filter paper and then dried etc., but those methods are requiring extra equipment/material and/or do require extra preparation.
At the bottom of this post you can find a list of a few papers on this topic that I found and read during my research, a simple web search
should get you the PDFs.


Aiming for maximum convenience in creating a long term storage medium I finally narrowed it down to the dehydrated grain spawn as this seemed
to be the most robust method to store a culture and most adequate for room temperature storage. There is no moisture/liquid and cooling
involved which I liked cause it's something new as everything else I got here that has mycelium in or on it is either slightly moist, dripping wet or at field
capacity and has to go in the fridge if I want to keep it around for longer.




The Method

The method I use to create dehydrated spawn is quite simple and goes along nicely with usual cultivation work.


STEP 1:

I transfer a good spoon full of spawn (50-80 grains) to a sterile petri dish without agar, the grain can be poured like when doing G2G. Other containers
might work as well as long as they are sterile but I think they should allow for lots of GE in order to dry out faster.


STEP 2:

The petri dish gets wrapped with micropore tape. I gave it a couple of layers and used some tension to avoid the tape from creasing too much and any
creases that formed were flattened out to make a good seal. The photo below is just a dummy dish for illustration purposes.



This protects the plate from getting contaminated during the time in the fridge.


STEP 3:

The dish gets placed in the fridge, best place is one of those covered compartments on the door.



As the cold air in the fridge has naturally a low rH the grains should dry out fairly quick. I do have one of those fridges that has an automatic defrostΒ and I don't know
how much of an influence that has on the drying time. I check on the grain every now and then and remove the plate when the spawn seems to have dried out.


STEP 4:

Once the grains are dry the plates are stored it in an air tight container. I used to put mine in vacuum sealer bags and sealed them without pulling a
full vacuum but I just put them in an airtight container now and replace the micropore tape on the plates with cling wrap.



Here are two plates of grains I prepared







Revival Test Part 1

I've prepared a few plates like this, 10 species in total. For my first test I used Oyster and Semilanceata cultures because I had plates
of these species in cold storage which I roughly made at the same time as the grains, so good to have a some sort of comparison.

The process is very simple just take a grain or two and put it onto an agar dish, just regular standard agar recipe, nothing special.

First Revival Test after 7/8 month in dry storage:

It took a week to show first signs of growth on the grains and I let it grow out for about a week before taking the photos so this is ~2 weeks after the transfer.
left: semilanceata after 7 month dry storage, right:pleurotus after 8 month dry storage




The control cultures from cold storage were doing a little bit better after ~2 weeks from transferring onto new plates.




Side by side after ~2 weeks




Side by side after ~3 weeks




Side by side after ~4 weeks




This definitely gives you something to work with, the new growth looks very clean. The dehydrated grain cultures are also catching up but I
managed to get some bacteria onto my cold stored Ps. semilanceata plate while taking a transfer. Will try these two cultures again in a years
time together with other cultures that I'm going to prepare in the meantime and I will update this thread with more results. If I manage
to revive 2 year old cultures I will start and fruit some of them to see how they are going to perform.





Revival Test Part 2

This is the second round reviving some of the dehydrated grain span I made. This time I am not dropping the kernels to agar but straight into
some LC broth.



The broth is 1% nutrient strength using LME and a drop of corn sirup. In this run I am reviving the following species:

Panaeolus cambodgeniensis, Psilocybe semilanceata, Pliscybe subaeruginosa,
Psilocybe cubensis (amazonian), Lentinula edodes 75 and Pleurotus pulmonarius

and here is the list of the samples and results. This list will be updated as results are coming in.

Grains dropped to LC on June 12 2021

Formula: 0



RESULTS:

The cultures PanCam "B" and PanCam "c" showed growth after 2 days they both were made 4 month ago.

This is PanCam "C" after 2 days and after 10 days, it came out clean when tested on agar and was put to grain showing good growth.




Spawn kernels of Pan cam I & Pan cam II have shown growth after 10 days in the broth.

This is Pan cam II after 10 days and I have put the LC to sterilized horse manure + straw substrate once it was full grown without testing on agar first.




After 4 weeks non of the other kernels showed any growth. I thought dropping the grain straight to LC would revive the cultures faster but maybe this
is not the way to go. I also didn't like that I was not able to verify the cleanliness of the culture without testing the LC on agar. This is why I
will keep dropping the grains to agar from now on as this will show me straight away whether the culture is clean or not unlike LC which leads me to
the next installment of this experiment.


Revival Test Part 3

As said above I am dropping grains to agar again. This time around I am testing the following species:

Panaeolus Cambodgeniensis (x3), Cyclocybe aegerita, Psilocybe papuana, Gymnopilus purpuratus, Psilocybe subaeruginosa,
Psilocybe semilanceata, Psilocybe cubensis, Lentinula edodes 75 and Pleurotus pulmonarius

and here is the list of the samples and results. This list will be updated as results are coming in.

Grains dropped to agar on July 5 2021

Formula: 1

It is not easy to determine when exactly new grows starts, on some kernels it seems that the first growth happens underneath the kernels hull/skin and
it takes a while to get through to the surface. Once the mycelium has broken through though the growth is quite vigorous.

The temperatures are currently a bit low here and I am not home much so all I can manage to do is to keep them at 58-68oF (14-18oC)
which makes for a relative slow growth rate.


RESULTS:

PanCam "B"

after 5, 9 and 15 days on agar




PanCam "C"

after 5, 9 and 15 days on agar




GymPur

after 5, 9, 15 and 22 days on agar




Papuana

after 15 and 22 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. The mycelium well outgrew the bacteria after 22 days making
it possible to easily transfer.

images after 8, 15 and 22 days




Conclusion for Test #3 so far:

This seems to work better than dropping kernels 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. This does indicate that 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. In the next round I will
focus on the cultures that haven't produced any growth so far and will play the numbers game by dropping ~10 kernels onto each plate.
In the meantime I will let the currently unproductive plates run for another 4 weeks.



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 a lot more grains on each dish to get a better chance of having viable
kernels on the plates. Each plate has between 30 and 50 kernels. I still got some more dried cultures that I haven't tested yet but will leave
that for sometime later.

Cultures tested this time are: Subaeruginosa from LME LC, Subaeruginosa from BRF LC, Semilanceata, Cubensis, Shiitake 75 and Phoenix Oyster.

Grains dropped to agar on August 11 2021: 

Formula: 2



RESULTS

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. Two more kernels showed growth after
20 days and the plate fully colonized eventually, second picture is a 1st transfer from the grain plate which shows clean growth and finally the grain plate
pinned after a couple more weeks.



All remaining plate were finally tossed after 35 days.


Final Conclusion

Well this wasn't the success I was hoping for but it wasn't a total failure either. I am sure that with a more refined drying method this
would work much better but I was aiming for the easiest way that was available to me at the time. If I would do this again I would dry the
spawn under controlled conditions in a climate chamber with a saturated salt that buffers the humidity level at around 10% so all grains dry
under the same conditions and will end up with the same moisture content. If I could I would also run tests with a freeze drying setup but
this will probably never happen. I'm focusing more on other long term storage methods now and currently I am putting together a collection of
my cultures as agar wedges in distilled water and LCs with a nutrient content of 0.05% which will be tested for viability in a years time.

This was a fun project, lets see how the other methods turn out.

:sun:


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

Titles of research papers on long term storage of mycelial cultures:
  • An example for comparison of storage methods of macro fungus cultures Schizophyllum commune

  • Evaluation of different storage methods for rapidand cost-effective preservation of Botryosphaeria species

  • Preservation of live cultures of basidiomycetes

  • Preserving cultures of wood-decaying Basidiomycotina using sterile distilled water

  • Saving-fungal-Cultures-Sterile-Water-By-Harris

  • Preservation of fungi in water (Castellani) 20 years

  • Preservation of fungal cultures of medical importance in distilled water




Edited by Baba Yaga (10/05/21 06:25 PM)


Extras: Filter Print Post Top
OfflineZakkery
I'm a teapot User Gallery


Registered: 10/15/18
Posts: 293
Loc: UK Flag
Last seen: 6 days, 14 hours
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests Part One [Re: Baba Yaga]
    #26943570 - 09/19/20 05:39 AM (3 years, 4 months ago)

Dope. Didn't know this was a thing. I've got plates just sat in my fridge but this is a nice long term option.

Do you think the grain would last a while in the fridge too? I don't have a vacuum sealer just yet.


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests Part One [Re: Zakkery]
    #26943580 - 09/19/20 05:52 AM (3 years, 4 months ago)

nah you wont need a vacuum sealer necessarily I think, as I said I don't pull a real vacuum in the bags its just to store it air tight like you
would store dried fruits. wrapping the dishes a few time with cling wrap and putting them in a drawer would be fine as well I suppose, but I just
started this long term experiment and will be curious to see what happens next year. My plan is to get away from cold storage so i will keep
everything at room temp.

And by the way i do this parallel to my usual plate/slant in the fridge storage. If I can revive 3 year old grains then I will call it good
enough to switch methods. 


Edited by Baba Yaga (09/19/20 06:09 AM)


Extras: Filter Print Post Top
OfflineForresterM
aspiring sociopath
Male User Gallery

Registered: 02/05/13
Posts: 9,351
Loc: Northeast USA Flag
Last seen: 24 days, 15 hours
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests Part One [Re: Baba Yaga]
    #26943613 - 09/19/20 06:25 AM (3 years, 4 months ago)

I've always thought dried mycelium would be good for storage, but never tested it.  Nice write up!

I think this would be an excellent method for tropical species like pink oysters that have trouble in the fridge as well.


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests Part One [Re: Forrester]
    #26944878 - 09/19/20 08:26 PM (3 years, 4 months ago)

Quote:

Forrester said:
I think this would be an excellent method for tropical species like pink oysters that have trouble in the fridge as well.




Yes I would try and use it on Pans as well, some do have a hard time bouncing back after a year in the fridge.


Extras: Filter Print Post Top
InvisiblePastywhyteMDiscord
Say hello to my little friend
Male User Gallery

Registered: 09/15/12
Posts: 37,808
Loc: Canada Flag
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #26952824 - 09/24/20 01:58 PM (3 years, 4 months ago)

This thread was moved from Mushroom Cultivation.

Reason:
Moved upon request of OP. I suggest it might be an idea to look into vacuum drying for the grains. Many people find that their fridges are damp, not dry. This may also be a more axenic friendly and a lot faster way to get the grains dry.


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Pastywhyte]
    #26953086 - 09/24/20 04:51 PM (3 years, 4 months ago)

Thanks Pasty for your input, have edited the OP.


Extras: Filter Print Post Top
InvisiblefahtsterM
Now With 33%More Faht
Male User Gallery


Registered: 06/17/06
Posts: 9,268
Trusted Cultivator
OG Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #26953557 - 09/24/20 10:10 PM (3 years, 4 months ago)

I use dried grains for storage as well.. I use special tyvek lids for drying the grain out.  I like the vacuum sealing.. gonna give that a go this next season.

Here’s the link to the lid set up https://www.shroomery.org/forums/showflat.php/Cat/0/Number/6886717/an/0/page/0

More procedure on extracting the glc later in the post.. might give ya some ideas

Faht


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: fahtster]
    #26953724 - 09/25/20 12:33 AM (3 years, 4 months ago)

Thanks Faht, great stuff! Definitely some inspiration there, will also look into Agar's thread but can't access it yet (no permission) have to wait a couple of days.

I thought about small jars with just bands and filter discs, but I don't have any discs atm, also no tyvek. I will look into
getting some tyvek suits for asbestos removal, they are pretty heavy duty or maybe get some cellulose filter sheets for sterile filtering
from a brewers shop or so, I might be able to get that stuff local, wanted to try some with my grain jars anyway.

My fridge is hard to beat though, it is so dry that it will dry out the
outer leaves on a white cabbage if left in there for a week and it will make a hand full of small fruits cracker dry in 3-4 days, that gave me the
initial idea to use it for drying my grain spawn. It's pretty empty most of the time though apart from a box of eggs and a block of cheese
and my slants and dishes lol.


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: fahtster]
    #26953750 - 09/25/20 01:06 AM (3 years, 4 months ago)

How long does it take for your myc-water to bounce back into action when
taken from dried grains?


Extras: Filter Print Post Top
InvisiblefahtsterM
Now With 33%More Faht
Male User Gallery


Registered: 06/17/06
Posts: 9,268
Trusted Cultivator
OG Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #26954046 - 09/25/20 08:18 AM (3 years, 4 months ago)

I’d say 2-3 more days than usual for the bounce back... myc is super resilient.

Oh yeah, the fridge is great for drying stuff out.  Lol, I actually have a little write-up about using the fridge to get cracker dry fruits in the summer when the humidity is high (it was before I had a dehydrator) ...now I just gotta remember where I put it.. here she is:

Quote:

I've posted this before but there are some new updates on it!! and they are good. lol ok, so when i've stated this before i said you should just stick the baggie in the fridge in the crisper drawer with the bag open and they would get about 10% drier in the summer when the RH  in the air is too much that your mushies don't dry completely no matter how long you have them under a fan. well... I was looking at this old crusty piece of pizza that i didn't cover in the fridge and it's rock hard after like three days... i can snap the thing in half. So i got the idea to use a breathable container to put them in the fridge with. i.e., a tyvek  mailer. yet another use for tyvek  from the P.O.!!  just put your partially dried mushies in a tyvek  mailer....



now just seal it up and toss it in your crisper! they will be cracker dry in probably less than a few days. some other benefits: it's sealed and you can't see whats in it... if someone saw it they probably wouldn't go ahead and break the seal on it. AND once you are done using it for drying, you can use the envelope to make self-healing jars lids or something. lol

fahtster




From this Thread

Great minds and all that :wink:. I will say that this last year I decided to store my grain jars in the fridge over the summer while I took a break to see if I could just start them up again in the fall (I use to store them at room temperature in a drawer.. I wanted to slow any growth down kinda like a slant)... it was a bad idea with my set up.. moisture definitely still got on the jars and made it’s way through my tyvek filters.. there was also glue residue on the filters from t tape I used to cover and make the lids.. it was moldy and made its way onto the grain.  That’s why I like your vacuum seal idea so much.. I’m going to use it to thwart the problem I had this summer.  Also gonna figure out a glueless route for the lids.

Faht


Edited by fahtster (09/25/20 08:41 AM)


Extras: Filter Print Post Top
InvisiblefahtsterM
Now With 33%More Faht
Male User Gallery


Registered: 06/17/06
Posts: 9,268
Trusted Cultivator
OG Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: fahtster]
    #27249296 - 03/12/21 03:40 AM (2 years, 10 months ago)

Just revived a 3 month old dried out clone on grain.. GLC’d the grains just like they were


Faht


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: fahtster]
    #27249305 - 03/12/21 03:49 AM (2 years, 10 months ago)

Noice! 
:highfive:


Extras: Filter Print Post Top
OfflineLukeTheDuke187
Stranger

Registered: 08/10/20
Posts: 163
Last seen: 6 hours, 10 minutes
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27270324 - 03/26/21 01:27 PM (2 years, 9 months ago)

Have you tried to dehydrate the grains before storage with a dehydrator maybe on a low temp setting?


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: LukeTheDuke187]
    #27270345 - 03/26/21 01:49 PM (2 years, 9 months ago)

Well you want to keep it sterile and I can't see how that could be achieved using a dehydrator. You could have the dishes sitting on a shelf
if you're in a reasonable dry climate but I would only put a few grains in each dish.


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27347354 - 06/13/21 10:47 PM (2 years, 7 months ago)

OK, here we go again.

Revival Test Part 2

This is the second round reviving some of the dehydrated grain span I made. This time I am not dropping the kernels to agar but straight into
some LC broth.



The broth is 1% nutrient strength using LME and a drop of corn sirup. In this run I am reviving the following species:

Panaeolus cambodgeniensis, Psilocybe semilanceata, Pliscybe subaeruginosa,
Psilocybe cubensis (amazonian), Lentinula edodes 75 and Phoenix oyster

Grains dropped to LC on 06/12/21

There is a table in the OP where I will update the results.


The pans are taking the lead with first signs of growth after only 2 days but the samples are much younger than the rest at 4 month old,
will be good to see if this is going to change once the pan samples are getting older. I hope this is not mold and the cultures are going
to be tested on agar of course.


First growth of the 4 month old pan cambo "C" on wheat kernels after only 2 days.



:sun:


Edited by Baba Yaga (07/13/21 09:05 PM)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27354847 - 06/19/21 06:42 AM (2 years, 7 months ago)

UPDATE 06/19/21:

Have put the two Pan LC cultures to agar today to check for cleanliness.

Still no growth on the other grains after a week, might be because the
jars are sitting at or below 60F or 15C. (Not home much lately so no
heating the house either.)

Will keep them a bit warmer, wait another week and report back.

Otherwise I'll try again on soft agar.


Edited by Baba Yaga (06/22/21 12:36 AM)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27358364 - 06/22/21 01:02 AM (2 years, 7 months ago)

UPDATE 06/22/21:

Spawn kernels of Pan cam I & Pan cam II have shown growth after 10 days in the broth.
There was nothing when I looked this morning but now there is clearly visible growth.


This is Pan cam II:



I really thought that the oyster would have poked out their head by now.

This would probably go a lot faster if kept at normal room temp so at the moment I am
not focusing on the speed cause this test was kept at relatively low temps.
There is growth is all that counts.


This is sample PC "C" after 10 days:



I have put some myc of this on agar but its not enough growth yet to snap a pic. I'm kinda tempted to just put it to a couple of grain jars.
I know, lazy right?!


Extras: Filter Print Post Top
Offlinegt40
I will proof smthng
Male User Gallery


Registered: 09/17/12
Posts: 502
Last seen: 8 days, 1 hour
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27359622 - 06/23/21 04:43 AM (2 years, 7 months ago)

Thanks a lot for sharing


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: gt40]
    #27386658 - 07/13/21 10:39 PM (2 years, 6 months ago)

Revival Test Part 3 is in progress now and has started on July 5th.

This time I am dropping grains to agar again. Have a look at the OP for the first update on this. I will ad further updates as results are coming in.


Edited by Baba Yaga (07/14/21 02:37 AM)


Extras: Filter Print Post Top
Invisiblejoze
Male


Registered: 11/10/20
Posts: 928
Loc: PNW
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:


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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.



Extras: Filter Print Post Top
OfflineSingularFusion
I'm a teapot User Gallery

Registered: 10/31/18
Posts: 1,208
Last seen: 7 hours, 45 minutes
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:


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


Extras: Filter Print Post Top
InvisibleGuerrilla
Bumbaclart

Registered: 01/30/21
Posts: 3,170
Loc: United Kingdom Flag
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!


--------------------
Being pissed on does not make you a real man.


Extras: Filter Print Post Top
InvisibleBlack and gifted
We are the Korg
 User Gallery


Registered: 08/24/10
Posts: 78
Loc: Alpha Centauri
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.


--------------------
"We can't win against obsession. They care, we don't.. They win."


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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)


Extras: Filter Print Post Top
InvisibleIchigo Kurosaki
Substitute Soul Reaper


Registered: 10/13/21
Posts: 150
Loc: Soul Society
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.


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


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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.


Extras: Filter Print Post Top
InvisibleIchigo Kurosaki
Substitute Soul Reaper


Registered: 10/13/21
Posts: 150
Loc: Soul Society
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:


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


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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.


Extras: Filter Print Post Top
Offlineyas champ
Stranger
 User Gallery
Registered: 06/27/19
Posts: 107
Last seen: 1 year, 1 month
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


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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%.


Extras: Filter Print Post Top
Offlineyas champ
Stranger
 User Gallery
Registered: 06/27/19
Posts: 107
Last seen: 1 year, 1 month
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:


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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.


Extras: Filter Print Post Top
Offlineyas champ
Stranger
 User Gallery
Registered: 06/27/19
Posts: 107
Last seen: 1 year, 1 month
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...) ?


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
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?


Extras: Filter Print Post Top
OfflineSingularFusion
I'm a teapot User Gallery

Registered: 10/31/18
Posts: 1,208
Last seen: 7 hours, 45 minutes
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


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


Extras: Filter Print Post Top
Offlineyas champ
Stranger
 User Gallery
Registered: 06/27/19
Posts: 107
Last seen: 1 year, 1 month
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: SingularFusion]
    #27543763 - 11/15/21 01:24 AM (2 years, 2 months ago)

I am in Algeria if you know (North Africa), I have already tried to grow the mycelium of agaricus bisporus, but it is always weak (a radius of 2 cm of mycelium only and the color tends to brown) and I wonder if the quality of the mushroom does not play a role in all this, would growing mycelium from a mushroom that has been transplanted several times (several generations) give the same result as growing it directly from the first bag of wheat?
I don't know if I succeeded in making you understand because my English is not good


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: yas champ]
    #27543780 - 11/15/21 02:52 AM (2 years, 2 months ago)

Ok, unfortunately I don't speak your language assuming that french is one of them but there are a few french members on here, I just can't
any names, sorry.

So you are trying to grow edible mushrooms for food? I have the feeling that this thread will not help you much as it is only about a few
experiments that didn't work that well.

If you are growing mushrooms for food then you might be better off asking this question in the gourmet & medicinal forum. Maybe make a
thread with a title like "Looking for someone who speaks french to help me with a few questions."

Here is the link to the gourmet forum:

https://www.shroomery.org/forums/postlist.php/Board/13

Good Luck


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: SingularFusion]
    #27543783 - 11/15/21 03:05 AM (2 years, 2 months ago)

Quote:

Nef said:
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






Hey Nef, thanks for popping by and sharing your experience, I always think of wood lover as the once that bounce back from dried up wood
substrates easily.

I am sure that this could work and drying it out to the right moisture content might be the key or freeze drying but all this would be too
much work for me. Maybe I will get back to this if I don't find an alternative.

Anyway, for long term storage I'm trying low nutrient LC out next and I got a few agar wedges in distilled water sitting in the cupboard for
almost a year now.

Will probably wait another year before testing anything.


Extras: Filter Print Post Top
Offlineyas champ
Stranger
 User Gallery
Registered: 06/27/19
Posts: 107
Last seen: 1 year, 1 month
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27543798 - 11/15/21 04:01 AM (2 years, 2 months ago)

I will search in the forum that you suggested, I may find an answer, thank you very much for the answers :rire:.
good luck.


Extras: Filter Print Post Top
OfflineSingularFusion
I'm a teapot User Gallery

Registered: 10/31/18
Posts: 1,208
Last seen: 7 hours, 45 minutes
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: yas champ]
    #27543856 - 11/15/21 06:15 AM (2 years, 2 months ago)

yeah for sure the wedge in distilled water also sounds like it might have some promise

good luck man :rockon:


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


Extras: Filter Print Post Top
Offlineshuna
 User Gallery

Registered: 03/07/21
Posts: 270
Last seen: 1 year, 4 months
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: SingularFusion]
    #27558548 - 11/26/21 03:22 PM (2 years, 2 months ago)

For your long term distilled water test, did you use a wedge of agar and what size, or did you scrape some mycelium off the agar surface?

I'm going to PC some 30ml tubes with say 75% tap water. Then drop say a 5x5mm wedge in and seal at room temp. Seems the easiest way to do it.

Scraping myc off the agar and getting it to stay in the water could be trickier, but then there will be less food for any potential contamination.

Edit: just tried dropping scraped myc to water and it drops easily.

Either way, looking forward to the results.

A grain jar in the fridge is meant to be a good method for long term storage, but bulky and less fridge the better.

:popcorn:


Edited by shuna (11/26/21 04:01 PM)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: shuna]
    #27558560 - 11/26/21 03:34 PM (2 years, 2 months ago)

Yes, a wedge of agar with a surface area of a stamp roughly.
Tap water should be fine I guess.

Grain in the fridge always turned after a few month in IME and as you
said it's bulky even if you use small jars and will be in the way if you
only have one fridge like me.

Another year and we will know.


Extras: Filter Print Post Top
InvisibleJustAPerson
Stranger
 User Gallery

Registered: 01/23/21
Posts: 22
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27568205 - 12/03/21 08:42 PM (2 years, 1 month ago)

Just mycelium in distilled water works great. I have a 3 year old culture that I fruited out just fine.


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: JustAPerson]
    #27568309 - 12/03/21 10:18 PM (2 years, 1 month ago)

Sounds awesome!

How are you transferring the mycelium?


Extras: Filter Print Post Top
InvisibleJustAPerson
Stranger
 User Gallery

Registered: 01/23/21
Posts: 22
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga]
    #27568685 - 12/04/21 07:14 AM (2 years, 1 month ago)

Cotton swab


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: JustAPerson]
    #27568696 - 12/04/21 07:30 AM (2 years, 1 month ago)

OK, would you care to explain the process a bit more please.

Swabbing a plate and then swirl the swab in distilled water? You do that more than once?

How much water per container and how much mycelium ends up in each?


Extras: Filter Print Post Top
InvisibleJustAPerson
Stranger
 User Gallery

Registered: 01/23/21
Posts: 22
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga] * 3
    #27568756 - 12/04/21 08:42 AM (2 years, 1 month ago)

It's a simple process.

1. Using one sterile cotton swab, swab one agar plate and pick up some mycelium
2. Put the swab in sterile water in one sterile culture tube
3. Shake the swab around until the mycelium comes off the swab and stays in the water.
4. Cap the one tube
5. Throw out your swab.
6. Repeat as desired with new tubes and swabs. Reuse the same plate if you want.

You need enough water to fill the culture tube. If you don't fill it all the way to the top you will have the chance that mycelium will float on top of the water and be exposed to air. Does that affect anything? I don't know! But if that does happen, you can try shaking the capped tube to get the mycelium into suspension.

I didn't measure the volume of mycelium per container, because that's hard. Much less than 1 mL, less than the volume of a single droplet of water. The amount of mycelium that gets stuck to a cotton swab. That's how much mycelium ends up per tube.

On the other side of the process

1. Put one sterile cotton swab in one centrifuge tube that has your water-suspended mycelium
2. Shake your swab around until you pick up some mycelium
3. Apply mycelium to agar
4. Toss your swab.
5. If there is mycelium left in your tube, save it, maybe? I've only done single uses.

There's also plenty of room to play with this. Double fist it and use multiple swabs on a single plate at once. Multiple swabs into multiple tubes simultaneously, if you have the dexterity.

Fundamentally, all you care about is getting mycelium into sterile water so that it is not exposed to air or food. However you get there.


Edited by JustAPerson (12/04/21 08:45 AM)


Extras: Filter Print Post Top
InvisibleBaba Yaga
β™₯ coir grower

Registered: 09/13/20
Posts: 3,957
Loc: Hyperspace Chicken Coop
Trusted Cultivator
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: JustAPerson]
    #27568984 - 12/04/21 11:46 AM (2 years, 1 month ago)

Thanks for writing this up in detail, using tubes that small is quite appealing. Will try this one day.


Extras: Filter Print Post Top
InvisibleJustAPerson
Stranger
 User Gallery

Registered: 01/23/21
Posts: 22
Re: Long Term Culture Storage On Dehydrated Grain + Revival Tests (moved) [Re: Baba Yaga] * 1
    #27569138 - 12/04/21 01:21 PM (2 years, 1 month ago)

I dug through my order history.

These 5 mL flat bottom tubes are what I used way back when. I selected them because they had a flat bottom. I don't recommend them, though, because they seal with a little o ring between the tube and the screw top, and that is finicky and not great.

Now I use these 15mL centrifuge tubeswith non-flat bottoms. Incrementally more annoying since they don't stand up on their own, but the screw top is way better with no o ring, and a stand is easily attainable or made.

You can also find 2 mL snap top centrifuge tubes. They may work for you, IDK.


Extras: Filter Print Post Top
Jump to top Pages: 1 | 2 | 3  [ show all ]

Shop: North Spore Bulk Substrate   Mushroom-Hut Liquid Cultures   Left Coast Kratom Buy Kratom Extract   Kraken Kratom Red Vein Kratom   Original Sensible Seeds Bulk Cannabis Seeds   PhytoExtractum Kratom Powder for Sale   Bridgetown Botanicals CBD Concentrates   Unfolding Nature Unfolding Nature: Being in the Implicate Order


Similar ThreadsPosterViewsRepliesLast post
* An Experiment In Long(ish) Term Culture Storage aural 1,066 2 08/15/02 10:44 AM
by DinoMyc
* Culture storage and agar plate alternative. SalviaEngland 1,834 2 01/18/03 05:48 PM
by ISH
* liquid culture to peroxyde grains...?! fredyjenkins 1,847 5 08/28/05 12:49 AM
by mycofile
* Dr. Rush wayne Storage tek. black_dove 2,708 5 03/23/02 03:10 PM
by Nighted
* Software for geneology and logging of cultures
( 1 2 all )
Eightball 6,246 24 07/22/05 04:58 AM
by Jpilot
* i want to grow enough to keep me set for life
( 1 2 3 all )
Dobie 5,001 41 11/14/02 05:44 AM
by Anonymous
* Storing cultures in mineral oil ?
( 1 2 all )
ragadinks 4,331 21 10/02/08 04:11 PM
by OZZ
* Agar mix? MeltingPenguin 1,537 4 09/29/01 09:44 PM
by Nyahbinghi

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: RogerRabbit, Pastywhyte, bodhisatta
4,583 topic views. 2 members, 5 guests and 0 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.046 seconds spending 0.006 seconds on 12 queries.