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OfflineGrzyby
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long term storage on filter paper
    #11491452 - 11/20/09 10:04 AM (11 years, 11 months ago)

Thought this was interesting. They said it works with bacteria and pathogenic fungi, maybe edibles and cubes too??

http://blog.mycology.cornell.edu/?p=383


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Offlinesolumvita
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Re: long term storage on filter paper [Re: Grzyby]
    #11492002 - 11/20/09 12:51 PM (11 years, 11 months ago)

Nice, I will certainly try it, on some of my cultures

Thanks Grzyby


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OfflineRogerRabbitM
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Re: long term storage on filter paper [Re: solumvita]
    #11494509 - 11/20/09 07:27 PM (11 years, 11 months ago)

I'm also going to be experimenting with this.  It could make culture slants obsolete if it works with edible fungi.  My use of wood in culture slants has a similar effect.  I've found if the culture has died in the test tube, I can pull the piece of wood out and lay it directly on a petri dish.  I've yet to lose a culture that had colonized the wood.
RR


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OfflineMycelio
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Re: long term storage on filter paper [Re: RogerRabbit]
    #11494839 - 11/20/09 08:22 PM (11 years, 11 months ago)

Drying definitely works for edibles. It is an old method for long term storage that just got forgotten during the past decades. I'm sure you can dry anything colonized, store without cooling, rehydrate and use years later.

Carsten


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InvisibleLokelYokel
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Re: long term storage on filter paper [Re: Mycelio]
    #11496701 - 11/21/09 03:19 AM (11 years, 11 months ago)

So you could rehydrate a square of filter paper, from Grzyby's link, with a drop of sterile water and then drop it into a jar of grain and then "Boom goes the Dynamite"?


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ReUse! RePurpose! ReCycle!  "Mom says use it again!"


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Offlinesolumvita
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Re: long term storage on filter paper [Re: LokelYokel]
    #11497112 - 11/21/09 05:58 AM (11 years, 11 months ago)

no it would be better to first go to agar to check for contams.


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OfflineFNFAL
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Re: long term storage on filter paper [Re: solumvita]
    #11497893 - 11/21/09 12:16 PM (11 years, 11 months ago)

If wood in a test tube facilitates long term storage, then wouldn't the wooden dowels used for log inoculation also work? If the dowel is colonized wouldn't you be able to put this into a ziploc and freeze?


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OfflineRogerRabbitM
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Re: long term storage on filter paper [Re: FNFAL]
    #11498215 - 11/21/09 01:32 PM (11 years, 11 months ago)

I would assume so.  If you have some colonized dowel pins, freeze a few for a month, and then thaw them out and let us know how they recover.  I know you can also freeze colonized rye grains for long term storage of many species.
RR


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OfflineFNFAL
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Re: long term storage on filter paper [Re: RogerRabbit]
    #11500759 - 11/21/09 09:08 PM (11 years, 11 months ago)

I've got some G. frondosa and a few species of oysters that have colonized some dowels. I'll freeze them and let you know.


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InvisibleDr Qi
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Re: long term storage on filter paper [Re: FNFAL]
    #11508279 - 11/23/09 01:54 AM (11 years, 11 months ago)

Hi; this is reinventing the wheel in a way. I don't know how good it would be for long term storage. But this technique was used a number of years back (50's/60's) by some to preserve specimens for mailing between colleges. This is not a well documented technique and the author may have felt he has rediscovered something not used though his readings. Which is not Plagiarism but a rediscovery of a technique lost if citation is give in time.  Dr Qi


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In life there are few words and witticisms that I may use when writing that have not been said or written in the past. So there is little that's unique to my writings. I do borrow and use ideas and information from published authors without making any claim that its original work done by myself. Just the best information that I have available at the time when writing, as the most I can do is to search for the truth and wisdom when writing.

May you learn to know yourself and live life happy, joyous and free; Dr Q'i


Edited by Dr Qi (11/23/09 02:49 AM)


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Invisiblenoobieshroomie
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Re: long term storage on filter paper [Re: RogerRabbit]
    #11514372 - 11/23/09 10:17 PM (11 years, 11 months ago)

Quote:

RogerRabbit said:
  I know you can also freeze colonized rye grains for long term storage of many species.
RR




thats pretty interesting
what are some of these species?

      -noobie-


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AMU

Best Thread Ever
CapZilla said:
not sure what GE and FAE are but i should probably get some.

Citric said:
Your signature is wrong on colonization temps!

GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT

ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef


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OfflineBrennus
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Re: long term storage on filter paper [Re: noobieshroomie]
    #11514974 - 11/23/09 11:36 PM (11 years, 11 months ago)

Quote:

noobieshroomie said:
Quote:

RogerRabbit said:
  I know you can also freeze colonized rye grains for long term storage of many species.
RR




thats pretty interesting
what are some of these species?

      -noobie-




Probably just about anything other than Straw Paddy mushroom mycelium. I've got a fully colonized grain jar of P. Cubensis F+ that managed to freeze solid in my lab fridge while I was moving into my new apartment. I think I'll do a G2G with it for shits and giggles and see if it works.


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Invisiblenoobieshroomie
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Re: long term storage on filter paper [Re: Brennus]
    #11517234 - 11/24/09 10:29 AM (11 years, 10 months ago)

sweet thanks
thats great to know:thumbup:

    -noobie-


--------------------
AMU

Best Thread Ever
CapZilla said:
not sure what GE and FAE are but i should probably get some.

Citric said:
Your signature is wrong on colonization temps!

GOOD JUDGMENT COMES FROM EXPERIENCE
EXPERIENCE COMES FROM BAD JUDGMENT

ROOM TEMP 70-75 IS BEST FOR COLONIZATION
Thank you mycochef


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InvisibleJavadog
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Re: long term storage on filter paper [Re: Grzyby]
    #13131738 - 09/01/10 01:22 AM (11 years, 1 month ago)

That article has this quote:

"Lactic Acid or Chloramphenicol can be added to any standard medium in order to reduce contamination by bacteria."

(this was when isolating a fungus, starting from a host sample)

Has anyone tried this?  I have a syringe to clean up and
this might be easier than a hot pour.

Thanks,

JD


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InvisibleMonkeyKnifeFight
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Re: long term storage on filter paper [Re: Javadog]
    #13134164 - 09/01/10 04:50 PM (11 years, 1 month ago)

Calocybe indica is another species that can't handle cold temperatures.  There are probably other tropical species.

This seems like a cool idea.  But instead of using master slants I have been storing my cultures using the distilled water method that has been talked about here many times.  It has many advantages for me.  I don't have to fill my fridge with tubes and also temperature sensitive species should be OK since the samples are stored at room temperature.  If a species can't handle the room temperature in my house year round it's probably gonna be a lost cause for me.

I've never read any downsides to using distilled water for storage.  Is there a compelling reason to use one of these other methods?


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OfflineFruitbuddy
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Re: long term storage on filter paper [Re: MonkeyKnifeFight]
    #13150774 - 09/05/10 03:15 PM (11 years, 1 month ago)

i would soak the sterilized filter paper in a nutritional solution, let it be overgrown and then dry it...but its a bit unecessary, you can just dry plates. this is the easiest way i can think of preserving sterile cultures.


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OfflineZOMan
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Re: long term storage on filter paper [Re: Fruitbuddy]
    #26047191 - 06/12/19 03:59 AM (2 years, 4 months ago)

I KNOW this is 8 years old,
That said I Have some lines that don't store well cold.
Seems this works by the sporulation of the target organism.
Has anyone got any updated info?


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OfflineParasoul
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Re: long term storage on filter paper [Re: ZOMan]
    #26404187 - 12/28/19 09:24 AM (1 year, 9 months ago)

Found this article when searching for ways of long term storage in distilled water - they use filter paper disks for transferring the mycelium from agar to sterilized distilled water :

Conservation of the mycelia of the medicinal mushroom Humphreya coffeata (Berk.) Stey. in sterile distilled water

I haven't tried it myself, but would love to hear some opinions.


Edited by Parasoul (12/29/19 03:20 AM)


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Re: long term storage on filter paper [Re: Parasoul]
    #26419853 - 01/06/20 09:12 PM (1 year, 9 months ago)

Since the thread is twice bumped, here's the state of the art: https://sci-hub.tw/10.1016/j.funbio.2013.12.002

Filter paper made the rounds in Fall 2019 as a good way to mail bacteria samples, which it is. This is often true for yeast and other molds so then people look at "fungi storage." We mostly care about mushroom producers (basidiomycetes) that are known to be more fragile than yeast and bacteria, i.e., you can't revive shiitake broth from minus 80 unless you make special preparations first.

The Homolka 2014 paper is divided into every known storage technique and it lists the most recently published storage length. Basidiomycetes are often underrepresented or else they die so grading techniques by "how many lived" is a bad idea. Note that freezing temps reflect the experimental designs described and are not constants. Any non-fluctuating freezer (no auto-defrost) is probably fine.

Subculturing (serial transfer) [agar slants]. Universally used despite their disadvantages. You can mitigate strain degradation with half-strength agar, a mineral or paraffin oil cover, or partially drying it.

Mineral or paraffin oil. Agar under mineral oil at minus 20 is good for our fungi. The linear growth of mycelium vs. budding of yeast and bacteria seems to require moisture for reliable storage. "Piaggio (1996) reported quite high viability of 127 fungal strains (mainly wood-rotting basidiomycetes) stored under oil and retrieved after 10y and 20y. A general survival rate was 80% after 10y and 93% of the surviving after 20y."

Storage in sterile water. It generally seems to work well for basidiomycetes provided the solution is isotonic (saline) and preferably not all aqueous. This is one of the first "solid teks" from 1939. My pet method, details at the end of the post.

Drying of fungal cultures. Drying them as on filter paper is good for spores, i.e., spore prints can last up to 5 years or more. For mycelium, you'd need silica gel to achieve worse results than water.

Freeze-drying (lyophilization). Large and often mold/bacteria libraries like ATCC freeze-dry their stuff in milk with a cryoprotectant. Not good for our goals: to reliably store a diverse library of hyphae so that we can easily revive a dead agar slant. "The effort to apply lyophilization to fungal hyphae in most cases has met with mixed success. Nevertheless, several results show that freeze-drying of basidiomycetes can be successful in some cases."

Cryopreservation. There are many methods that range from minus 80 to liquid nitrogen on a variety of substrates. Homolka's own pet method (a good one except for working with perlite) involves growing a 10% glycerol broth with sterile perlite crystals, then draining the broth and storing the crystals. This section is the bulk of the paper and it's worth reading. I want to focus on techniques you can do with a freezer that doesn't auto-defrost.

Based on all this info, I think there are a few criteria that stand out as a rule of thumb: "basidiomycetes keep best either in cryo conditions or those closest to their natural habitat." Well, where do we usually find the best mushrooms? In places that are wet but not underwater, gritty but not salty, cold but not permafrost, where air and light are either benign or helpful.

My experiments have settled on some good tools. Wash agar from a clean plate in 0.6% sea salt saline to suspend the mycelium, and mix it well, 50:50 with pure glycerol. Inject this into BD Vacutainer serum tubes (red top), which auto-fill exactly to 10 mL and leave a substantial air plenum. Store the tubes in a styrofoam box in the freezer. The 0.6% sea salt is for metabolic purposes: to have 33% less salt (water shouldn't be a limiting factor) and to randomize the salt contents (trace minerals may become relevant).

A larger strategy would have multiple time scales, including decades if you can afford cryo for that long. The simplest would be two scales: agar slants to make plates from, and frozen glycerol/saline to have reference backups. Trading very low metabolism samples and high quality genetic data would be like downloading a cool file and a crypto signature to verify it. :loveeyes:


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Edited by AndyHinton (01/06/20 09:21 PM)


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