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Invisiblevinsue
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clone from dried caps
    #2390329 - 03/01/04 06:04 AM (19 years, 11 months ago)

Is it possible to start a clone on agar using mushroom sample that was dried and has been frozen for over 1 year? Original sample was extremely potent. Thanks


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OfflineFloyd_
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Re: clone from dried caps [Re: vinsue]
    #2390396 - 03/01/04 07:16 AM (19 years, 11 months ago)

unless its a species that can withstand freezing its most assuredly 100% dead. And even if it is a species that can withstand freezing..its still probably dead


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Anonymous

Re: clone from dried caps [Re: Floyd_]
    #2390667 - 03/01/04 09:37 AM (19 years, 11 months ago)

No clone but you can maybe get some spores to germinate.


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Offlineabhi
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Re: clone from dried caps [Re: vinsue]
    #2406436 - 03/07/04 08:30 PM (19 years, 11 months ago)

I wouldn't bet on it, but if you have the resources to spare you may as well try it. You could get lucky.


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OfflineScourge
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Re: clone from dried caps [Re: ]
    #2407038 - 03/08/04 12:40 PM (19 years, 11 months ago)

i agree, spore germination would probably be your best bet.


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OfflineMitchnast
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Re: clone from dried caps [Re: Scourge]
    #2424848 - 03/12/04 04:19 PM (19 years, 11 months ago)

i bet if you tried to clone a dried cap, you would germinate the spores anyway, unless you used peroxide first


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OfflineRogerRabbitM
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Re: clone from dried caps [Re: vinsue]
    #2438551 - 03/16/04 08:47 AM (19 years, 10 months ago)

Dried caps have been known to start growing again on agar. Go for it.


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Anonymous

Re: clone from dried caps [Re: RogerRabbit]
    #2438564 - 03/16/04 08:55 AM (19 years, 10 months ago)

What!?!? Thats crazy are you just fucking around or serious. Well any way thats cool!


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OfflineCaptainFuture
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Re: clone from dried caps [Re: RogerRabbit]
    #2440093 - 03/16/04 03:53 PM (19 years, 10 months ago)

NEVER!
A dried cap is not a sklerotia...


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Offlinesnowspeed1
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Re: clone from dried caps [Re: vinsue]
    #2441863 - 03/17/04 01:06 AM (19 years, 10 months ago)

Actually i read a post on this board about a guy who did it with a 20 year old mushroom. there was little left but dust. never underestimate the power of a determined nature. he used peroxide on it as well as a safety measure.


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InvisibleJoshua
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Re: clone from dried caps [Re: snowspeed1]
    #2441891 - 03/17/04 01:19 AM (19 years, 10 months ago)

"unless its a species that can withstand freezing its most assuredly 100% dead."

This is true.

Cubensis fruit bodies do not survive dessication. I personally am not aware of any species whos fruitbody will survive dessication, especially for 20 years.

It is probable that what was grown from the cap were spores and not tissue.

Joshua


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OfflineRogerRabbitM
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Re: clone from dried caps [Re: vinsue]
    #2442524 - 03/17/04 08:24 AM (19 years, 10 months ago)

With all due respect to those who disagree or have never done it, you can take dried mycelium and get it growing again on agar. I've done this many times with dry mushrooms. The stipe works better than the cap. This is not due to spores germinating, but due to the dried mushroom tissue rehydrating and beginning to grow again.


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Offlineragadinks
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Re: clone from dried caps [Re: RogerRabbit]
    #2442632 - 03/17/04 08:57 AM (19 years, 10 months ago)

Does that depend on the species? With what species did you try that out?
Did you just put them on agar or did you rehydrate them first?


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OfflineNewdude1
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Re: clone from dried caps [Re: RogerRabbit]
    #2442642 - 03/17/04 08:59 AM (19 years, 10 months ago)

Seems to be some debate on this, I think maybe it calls for an experiment with pictures RodgerRabbit? mabye some dried myc, dried fruit, and dried cap spores, that would be really cool.


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OfflineShroomInhaler
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Re: clone from dried caps [Re: Newdude1]
    #2443106 - 03/17/04 11:43 AM (19 years, 10 months ago)

So what some of you are saying is that if i purshase dry mushrooms from someone, I don't need a spore syringe or anything, I can just try and use the dried msuhrooms??


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OfflinePooPs
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Re: clone from dried caps [Re: ShroomInhaler]
    #2443157 - 03/17/04 11:59 AM (19 years, 10 months ago)

I'll bet there were spores on the stem, causing germination of more mycelium..

But that's just my dumb ass opinion !


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InvisibleJoshua
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Re: clone from dried caps [Re: RogerRabbit]
    #2443463 - 03/17/04 01:16 PM (19 years, 10 months ago)

This is news to me! I'll try it and see what happens.

Joshua


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Anonymous

Re: clone from dried caps [Re: Joshua]
    #2443712 - 03/17/04 02:09 PM (19 years, 10 months ago)

Ok well though I am skeptic. I do believe that this could be true. Yeast a cusin of fungi, does reanimate when dried and rehydrated. So I guess this could be true for mushrooms. As long as the cell walls arnt damaged in the drying process. Hmmmm


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InvisibleJoshua
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Re: clone from dried caps [Re: ]
    #2447599 - 03/18/04 02:43 PM (19 years, 10 months ago)

Surely someone else has knowledge on the subject!

Joshua


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Great books for inquiring minds!

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Offlinebobmarley4prez
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Re: clone from dried caps [Re: ]
    #2447614 - 03/18/04 02:48 PM (19 years, 10 months ago)

Yeast a cusin of fungi

yeast is a term used to describe a unicellular fungi of three of the four phyla of fungi (zygomycota, ascomycota, and basidiomycota(most mushrooms).
Peace


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Offlineragadinks
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Re: clone from dried caps [Re: bobmarley4prez]
    #2447660 - 03/18/04 03:00 PM (19 years, 10 months ago)

That means that yeast is a fungi ?


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InvisibleJoshua
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Re: clone from dried caps [Re: ragadinks]
    #2448070 - 03/18/04 04:12 PM (19 years, 10 months ago)

Yes.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


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InvisibleYarry
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Re: clone from dried caps [Re: bobmarley4prez]
    #2448076 - 03/18/04 04:14 PM (19 years, 10 months ago)

i bet if you rehydrated them they would grow, i think the problem here is that they were frozen. From what i kno freezing will damage cell walls and other important cell structures, so i doubt they will grow.

Joshua, you should try growing from dried shrooms to see if it works. If anyone could get it to work youd be the man


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InvisibleJoshua
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Re: clone from dried caps [Re: Yarry]
    #2448147 - 03/18/04 04:26 PM (19 years, 10 months ago)

RogerRabbit says he has accomplished this, I guess he might be the man. Although I did see a pic of him wearing a bra and bunny ears once :lol:

I will give it a try.

Joshua


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Great books for inquiring minds!

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InvisibleMilkVein
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Re: clone from dried caps [Re: Joshua]
    #2448159 - 03/18/04 04:30 PM (19 years, 10 months ago)

lol, i saw that pic too^


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Offlinesnowspeed1
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Re: clone from dried caps [Re: vinsue]
    #2450558 - 03/19/04 03:15 AM (19 years, 10 months ago)

Freezing destroys cell walls because the water inside the cell expands 10% when frozen. just like sticking a cheap bottle of beer in the freezer and having it explode. if the shrooms were dried then their would be no danger of this.
and rogerrabbit is definitely right about the shroom itself regrowing.
for instance, you can pick up a mushroom at your local supermarket and start it growing on medium. you know that its been picked for quite some time, yet it still grows.


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InvisibleJoshua
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Re: clone from dried caps [Re: snowspeed1]
    #2450636 - 03/19/04 04:25 AM (19 years, 10 months ago)

Your reasoning for freezing is true.

There is a very big difference between a fresh store bought mushroom and a mushroom that has been desiccated.

Joshua


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Great books for inquiring minds!

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Offlineblackout
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Re: clone from dried caps [Re: Joshua]
    #2451114 - 03/19/04 09:11 AM (19 years, 10 months ago)

Frozen sclerotia is meant to be "clonable". i am very surprised nobody has experimented with this or other storage methods. has anybody frozen syringes and found they still worked?
if you cut a sterile segment of mushroom and put it in a sterile jar how long do you think it would last in the fridge?


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InvisibleSpeeker

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Re: clone from dried caps [Re: blackout]
    #2451225 - 03/19/04 10:10 AM (19 years, 10 months ago)

I have read somewhere that some dried mycelium preparations are
viable about six months...

don't remember if they where freeze-dried , though...

dried mycelium preparations


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InvisibleJoshua
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Re: clone from dried caps [Re: blackout]
    #2451644 - 03/19/04 01:21 PM (19 years, 10 months ago)

I don't doubt sclerotia, stomata, or spore survival as the structures are designed for surviving environmental extremes.

http://bugs.bio.usyd.edu.au/Mycology/sandf/sclerotia.htm

I will try revitalizing dried fruitbody flesh. I think it would be cool if some other users tried the same independantly and reported the results within this thread.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


Edited by Joshua (03/19/04 01:23 PM)


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Anonymous

Re: clone from dried caps [Re: Joshua]
    #2451799 - 03/19/04 02:09 PM (19 years, 10 months ago)

So should I take some dried mycelium from an old plare and transfer it? Thats all I have to play with.


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InvisibleJoshua
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Re: clone from dried caps [Re: ]
    #2452107 - 03/19/04 04:05 PM (19 years, 10 months ago)

What's a plare?

I got it, a plate? I think if anything you should try the mycelium from the fruit body. Dense mycelium (such as occurs in sclerotia, stromata, and fruits) is more apt to survive than the loose colonizing mycelium.

Joshua


Edited by Joshua (03/19/04 04:25 PM)


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Offlinelittlejohn747
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Re: clone from dried caps [Re: Joshua]
    #2452155 - 03/19/04 04:18 PM (19 years, 10 months ago)

just to throw somthing into the mix i have used 4 frozen spore syringes. and they did grow out. the spores were 4 different strains to.


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OfflineRogerRabbitM
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Re: clone from dried caps [Re: vinsue]
    #2452227 - 03/19/04 04:41 PM (19 years, 10 months ago)

Prepare the agar a bit on the wet side to help hydrate the tissue. Also, be ready with fresh plates to transfer into, because all the contamination on the dried fruit will also begin to grow. You'll have to transfer healthy mycelia as soon as you see it to a new petri dish. It generally takes ten days to two weeks, sometimes longer, for the dried fruit to begin to grow again. Unfortunately the contamination doesn't need that long, so you have a bit of work to do to clean up the culture, but it will work.


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"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Offlinethisspock
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Re: clone from dried caps [Re: bobmarley4prez]
    #2548976 - 04/11/04 05:25 PM (19 years, 10 months ago)

A Classification of the Fungi:

Division 1 Gymnomycota - 'Naked' no cell walls contians slime molds
Class 1. Acrasiomycetes
2. Myxomycetes
Division 2 Mastigomycota - flagellated
Class 1. Chytidiomycetes- single cell 1 flagella
2. Oomycetes - biflagellated
Division 3. Amastigomycota - no flagella
Class 1 Zygomycetes - breadmolds
2. Ascomycetes - -Yeast fits in here- sac fungi
3. Basidiomycetes - club fungi, ie. mushrooms.
4. Deuteromycetes - no identified sexual phase.

Yes yeast and mushrooms are located in the same division of fungi, but it only classifies them as 'true' fungi; saprobic, stores in the form of glycogen, chitin cell wall, no flagella and sexual and asexual reproduction.

Yet a mushroom cap and a mushroom stem are NOT sclerotia and therefore are not tissue designed to deal with harsh conditions. A typical Basidiomycota sclerotia is usually derived from the mycellium stage and thus caps and stems cannot be rehydreated. A dead fruting body is simply chitin and leftover organic material with no means of division or reproduction even after hydration. To have a agar plate start growth from a dried piece of mushroom means that a spore has germinated on it.

I am fairly sure the same is true with yeast. A dead yeast cannot divide any longer but yeast ascospores can remain viable for long periods of times.

*I think* a yeast forms a sclerotia, and it can rehyrdate and grow. since a yeast is a single cell fungi the sclerotia will be the yeast in its entirety. However just being in the same division doesn't mean a P.cubensis sclerotia is of teh same properties.



~~Please correct any mistakes I may have made~~~


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OfflineRogerRabbitM
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Re: clone from dried caps [Re: thisspock]
    #2553707 - 04/13/04 08:29 AM (19 years, 10 months ago)

"To have a agar plate start growth from a dried piece of mushroom means that a spore has germinated on it".

Not so. You can even take a piece of the inside of a dried stem and get it to grow again on agar. No way a spore could be on the inside of the stem where it's protected. Dried mycelium of many species can be hydrated and coaxed to grow again. You are correct though. . .A dried cap or stem is not sclerotia. Cubies don't form sclerotia. However, a dried cubie can be rehydrated and grown out again on agar. Try it. You'll see the growth coming in many directions from the piece of stem you place in the agar. You will have no doubt that you have 'cloned' that piece if you'll try it.


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InvisibleJoshua
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Re: clone from dried caps [Re: RogerRabbit]
    #2553786 - 04/13/04 09:02 AM (19 years, 10 months ago)

I will test this very soon, just waiting on new plates.

Joshua


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The Shroomery Bookstore

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InvisibleSpeeker

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Re: clone from dried caps [Re: Speeker]
    #2553854 - 04/13/04 09:31 AM (19 years, 10 months ago)

In previous post I used clumsy term "dried mycelium preparations" that I took from that webpage. It is these "dried mushroom spawn" packets/bags that I was referring to. Same thing probably?

http://www.seedsofitaly.sagenet.co.uk/funghi.htm
http://www.cjindustries.co.uk/cg020001.html
micelio secco .... Italian sites...  :confused: don't understand Italian:
http://www.funghimara.it/fai_da_te.php?#micelio

http://www.funghiitaliani.it/rivista/main/la%20coltivazione%20dei%20funghi.htm
http://www.xmx.it/funghi.htm


Anybody knows how they make these preparations?
Are they just dried or perhaps freeze-dried?

perhaps somebody understands enough Italian to find this out... :grin:



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Offlinethisspock
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Re: clone from dried caps [Re: RogerRabbit]
    #2556399 - 04/13/04 06:44 PM (19 years, 10 months ago)

I was under the impression that in basidiocarps sclerotia was formed on the mycellium level.


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InvisibleJoshua
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Re: clone from dried caps [Re: thisspock]
    #2557756 - 04/14/04 01:30 AM (19 years, 10 months ago)

It's all mycelium, except for spores. The different morphology that the mycelium takes forms the different structures such as carpophores, sclerotia, stroma and vegetative growth. The colonizing mycelium is of the vegetative type, I imagine this is what you are referring to as the, "mycellium level."

Sclerotia form at or below the level of the colonizing substrate.

Joshua


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The Shroomery Bookstore

Great books for inquiring minds!

"Life After Death is Saprophytic!"


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InvisiblePinback
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Re: clone from dried caps [Re: RogerRabbit]
    #2631384 - 05/03/04 10:18 AM (19 years, 9 months ago)

I've tried this with dried agar. 20 days ago, I cut a couple of wedges from a colonized cubensis plate and put them in an empty plastic petri dish, which was stored at room temperature. After laying there and drying for 19 days, I placed one of them on a fresh non-peroxidated PDYA plate. The wedge looked really dry to me and was very hard. To my surprise, as I did not expect this result, there is growth today, only 1.5 days after I put it there:



I plan on trying the other wedge in 30 days if I remember it.


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Anonymous

Re: clone from dried caps [Re: Pinback]
    #2631392 - 05/03/04 10:22 AM (19 years, 9 months ago)

Yeah man, You Rock!


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Anonymous

Re: clone from dried caps [Re: Pinback]
    #2631537 - 05/03/04 11:20 AM (19 years, 9 months ago)

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InvisiblePinback
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Re: clone from dried caps [Re: ]
    #2632019 - 05/03/04 01:37 PM (19 years, 9 months ago)

Thank you both for your comments! However, I only repeated what RogerRabbit did, but with agar instead of mushroom tissue to minimize the risk of having spores germinating.


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Re: clone from dried caps [Re: Pinback]
    #2632169 - 05/03/04 02:09 PM (19 years, 9 months ago)

Thank you very much for posting the results! Now is as good of a time as any to transfer a dried peice of stem I've prepared for this experiment to agar. Results to follow.

Please post results of your plate as the growth proceeds. I think it's important to eliminate the growth as some form of contaminant mold.

Joshua


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OfflineGr0wer
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Re: clone from dried caps [Re: Joshua]
    #2632483 - 05/03/04 03:04 PM (19 years, 9 months ago)

Are you sure its not a contam?


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Anonymous

Re: clone from dried caps [Re: Joshua]
    #2633454 - 05/03/04 06:01 PM (19 years, 9 months ago)

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InvisibleJoshua
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Re: clone from dried caps [Re: ]
    #2634374 - 05/03/04 08:58 PM (19 years, 9 months ago)

There are vendors that sell dehydrated spawn. I don't have a link yet....

Anno provided me with a gourmet link. He said he tried the product without success.

I'll definately provide my results when they occur.

Joshua


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Anonymous

Re: clone from dried caps [Re: Joshua]
    #2634399 - 05/03/04 09:02 PM (19 years, 9 months ago)

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InvisiblePinback
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Re: clone from dried caps [Re: Joshua]
    #2641978 - 05/05/04 02:24 PM (19 years, 9 months ago)

It definitely looks like cubensis mycelium:


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Re: clone from dried caps [Re: Pinback]
    #2644971 - 05/06/04 10:44 AM (19 years, 9 months ago)

Nice work! I've also done this several times, but didn't do it for this thread, to allow someone else to. It's always better when an independent researcher confirms one's work. I'm anxious to see Joshua's results as well.

Five mushies for mr. pinback!


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Re: clone from dried caps [Re: Pinback]
    #2646539 - 05/06/04 04:25 PM (19 years, 9 months ago)

So is that a wedge of a mushroom cap?
Just to play devil's advocate:
How did you make sure that no spores were present when you put it into the agar. I Just cant see a way to determine that you have a spore free sample without a microscope.

This is a really neat experiment though, I can't way to see how Joshua's experiment turns out too.


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Re: clone from dried caps [Re: thisspock]
    #2647227 - 05/06/04 07:15 PM (19 years, 9 months ago)

Quote:

I've tried this with dried agar. 20 days ago, I cut a couple of wedges from a colonized cubensis plate and put them in an empty plastic petri dish




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Re: clone from dried caps [Re: Joshua]
    #2692689 - 05/17/04 06:31 PM (19 years, 8 months ago)

I placed a desicated stem section onto agar plates. The section was first allowed to sit in a 3% H2O2 solution for about 2 minutes. The stem was then rinsed with distilled water.

The stem was looked at after 5 days, nothing but bacterial contamination.

As the stem contaminated, my proceedure was not sterile and therefor led to inconclusive results.

I'll have to try again.

Joshua


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Re: clone from dried caps [Re: Joshua]
    #2710239 - 05/21/04 07:37 AM (19 years, 8 months ago)

QUITE the post!!!

Been gone a while folks, but this caught my eye i think SPORES grew?
BUT if it was tissue im AMAZED MAN....i would try it but always get contamed plates.


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Re: clone from dried caps [Re: bluelou]
    #2718447 - 05/23/04 01:21 PM (19 years, 8 months ago)

Guys,
This is old science. It's been done for many years in the commercial mushroom industry. It's the standard way they keep and trade isolated strains of edible mushrooms. They dessicant dry the fruitbodies, then powder them. They store the powder until ready for use, then sprinkle it on agar to regenerate the strain. It stops senescence in its tracks.

Try it without soaking in peroxide first. That only weakens the mushroom myc so it doesn't take off as fast. Watch for the first signs of myc growth, and immediately transfer that to a new petri dish. Do this a time or two and you'll have a clean specimen, even from years old dried fruits. It works, trust me.


Edited by RogerRabbit (05/23/04 01:22 PM)


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Re: clone from dried caps [Re: RogerRabbit]
    #2718955 - 05/23/04 03:34 PM (19 years, 8 months ago)

yeah, i've seen other reports of similar. old doc charlie at drooldonkey reported that kernels of grain covered with mycellia, left at the bottom of spawn jars for months and very dry, grew out again once re-hydrated.
and those kernels were never anywhere near any spores
rodger's just taken the next step.


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Re: clone from dried caps [Re: RogerRabbit]
    #2738175 - 05/27/04 06:46 PM (19 years, 8 months ago)

Quote:

This is old science.




That must be my problem...it's an age thing;)

Joshua


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Re: clone from dried caps [Re: Joshua]
    #2814540 - 06/21/04 03:38 PM (19 years, 7 months ago)

About a week ago I mixed up some PDYA with kanamycin (an antibiotic). Spilled a small droplet onto agar, and then took a huge stem from some really strong stuff on the street, must have been from a bulk grow, took a fluffy part from the inside and put it on the agar. Now some mycelium is growing wonderfully, no contams grew. Although I'm not 100 percent sure its cubensis mycelium im pretty sure it is. I'll take a picture and upload it soon.


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Re: clone from dried caps [Re: sci33]
    #2814949 - 06/21/04 06:15 PM (19 years, 7 months ago)



And a shot from underneath the plate, the brown area is the fluffy inner stem I placed on the agar. The mycelium is growing very thick mostly because I just had the nutrients of this agar batch a bit on the high side by accident. I will probably mix up another batch of agar/petris and do a second isolation to make sure no contams grew beneath the mycelium.

I will take some shots of the mushies this came from, and then maybe start a grow log for this unknown 'commercial' strain, start to finish.





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Re: clone from dried caps [Re: sci33] * 1
    #2815153 - 06/21/04 07:31 PM (19 years, 7 months ago)

> Spilled a small droplet onto agar, and ...

You spilled what on agar - the antibiotics or the PDYA ?


Anyway, great experiment  :thumbup:


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Invisiblesci33
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Re: clone from dried caps [Re: ragadinks]
    #2815351 - 06/21/04 09:13 PM (19 years, 7 months ago)

oops that meant to say spilled a small droplet of agar onto the petri, as seen in the pic.


--------------------
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Re: clone from dried caps [Re: sci33]
    #2816272 - 06/22/04 06:00 AM (19 years, 7 months ago)

Oh, now I know what I see in the pic :smirk:
Was a little bit confused by the descritpion.


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Re: clone from dried caps [Re: sci33]
    #2816575 - 06/22/04 08:34 AM (19 years, 7 months ago)

Very cool! So you don't know anything about the age of the dried fruitbody?
Great job regardless, I like the agar drop idea!


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Re: clone from dried caps [Re: Suntzu]
    #2816825 - 06/22/04 10:55 AM (19 years, 7 months ago)

Age of the dried fruitbody.. not sure, but I know it came from a large bulk grow since the person had a LARGE amount and they were all huge and potent (probably dung based substrate). I took the base of the stem in the flowhood, broke it open, and there was some fluff in there, took it with the scalpel and placed it on the agar.. no sterile procedure at all other than working in the flowhood. The agar itself was a bit on the thick side, and the reason I only placed a drop was that I knew it would take a while to reanimate the mycelium, and so I didn't want to risk contams growing on the outer surface of the agar while the middle is still colonizing. I'm sure the Kanamycin must have helped as well to prevent any bacteria. Will keep everyone updated on the progress.


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Re: clone from dried caps [Re: sci33]
    #2817632 - 06/22/04 03:38 PM (19 years, 7 months ago)

Very excellent!!!

Joshua


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Re: clone from dried caps [Re: bobmarley4prez]
    #2818156 - 06/22/04 05:45 PM (19 years, 7 months ago)

Hey what species can withstand below freezing temperatures? I actually have tried this before with a dried mushroom and it got contaminated. I just said screw it and stuck with syringes and mycelium.


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Re: clone from dried caps [Re: Philanthropist]
    #2857014 - 07/04/04 07:27 PM (19 years, 7 months ago)

I'm really skepticle about this whole thread. Stamets in TMC (in the tissue culturing chapter -- sorry, don't have the book) said that *fresh* (non-dried) fruitbodies need to be less than 48 hours old to clone effectively.

Any luck, Joshua? Could someone, at least, back this up with some literature? I think it's a lot more likely that contaminant mould or mycelium from a few spores would grow than from dead mycelium. I don't think dried mycelium would be able to carry out the basic cellular functions needed to survive.

If this is, in fact, possible, then why did Stamets say that in TMC?

Thx,

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Re: clone from dried caps [Re: micro]
    #2858390 - 07/05/04 09:28 AM (19 years, 7 months ago)

I was skeptical too, and am still a bit, but it has worked for me (from agar) twice. Now I have a colonized jar of grain from a piece of agar that had been dried for 55 days. Please try and repeat it, it's really a minimal amount of work.

I did a quick PubMed search but didn't find any entrys on room temperature drying. There were however at least one entry on lyophilization: Functional integrity and structural stability of freeze-dried ectomycorrhizal fungi established through viability assays. Some culture banks also sell lyophilized cultures. Source. There are of course differences between lyophilized and RT dried, but dried mycelium does however revive.


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Re: clone from dried caps [Re: micro]
    #2858910 - 07/05/04 01:35 PM (19 years, 7 months ago)

Quote:

micro said:
I'm really skepticle about this whole thread. Stamets in TMC (in the tissue culturing chapter -- sorry, don't have the book) said that *fresh* (non-dried) fruitbodies need to be less than 48 hours old to clone effectively.

Any luck, Joshua? Could someone, at least, back this up with some literature? I think it's a lot more likely that contaminant mould or mycelium from a few spores would grow than from dead mycelium. I don't think dried mycelium would be able to carry out the basic cellular functions needed to survive.

If this is, in fact, possible, then why did Stamets say that in TMC?

Thx,

--
Micro




He didn't.
In fact it was Stamets who first told me you could clone a totally dry fruit when I found a dry one that had been out in the sun for months. He was right, and it worked.


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Re: clone from dried caps [Re: RogerRabbit]
    #2859676 - 07/05/04 07:31 PM (19 years, 7 months ago)


You people are funny >>> doubting RR is a bad bet.

The not being able to freeze mycelium et al, and have it recover is also a myth.


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Re: clone from dried caps [Re: charlieXxXxX]
    #2859877 - 07/05/04 09:11 PM (19 years, 7 months ago)

My agar sample was transfered to new plates, and jars, which are almost done colonizing. Will keep updating.


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Re: clone from dried caps [Re: charlieXxXxX]
    #2861257 - 07/06/04 11:26 AM (19 years, 7 months ago)

Quote:

You people are funny >>> doubting RR is a bad bet.




It's my reson d'etre to doubt everything -- nothing personal. I just want either something from literature or the experiment repeated a few times. Maybe I'll go and buy some dried shiitakes from the store and try them out....

Just sounded strange -- that's all.

I'm also like 90% sure it said that somewhere in TMC -- I think under cloning a fruitbody -- but I don't have the book....

Peace!

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Re: clone from dried caps [Re: micro]
    #2861310 - 07/06/04 11:56 AM (19 years, 7 months ago)

Quote:

micro said:
I'm really skepticle about this whole thread. Stamets in TMC (in the tissue culturing chapter -- sorry, don't have the book) said that *fresh* (non-dried) fruitbodies need to be less than 48 hours old to clone effectively.
Micro




You're correct. Stamets says in TMC (and GGMM) that it becomes
difficult to isolate a pure culture if the specimens are too old.

But he does not say that it can't be done...


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Re: clone from dried caps [Re: Speeker]
    #2901995 - 07/18/04 09:39 AM (19 years, 6 months ago)

Dried or fresh the molecular degradation starts to take place immediately making a sample unviable or at the very best myc with little vigor over time. I have tried to revive myc on plates that have dried up over time with no success. That fluff from the inner stem sounds like it was still very much viable and most likely not very old at all so i'm not skeptical at all. This is not a technique for long term storage at all and it would be foolish to think otherwise IMHO. Someone turned me on to this thread with hopes of long term storage and now I know he took this out of context. The distilled water tek has caught my interest over myc under mineral oil in slants at the moment for long term. I respect RR alot, I know the commercial culture storage facilities still use deep freezing in liquid nitrogen to stop degradation through molecular activity (fact) also powderizing a mushroom would have to be at it's earlier stages in order for spores to not be a concern. I have friends that work in the commercial mushroom industry and I will be inquiring as to there procedure and their thoughts on what the lifespan of the powderized product would be. For something not to oxidize or degrade over a period of time without a stop in molecular activity is new to me. It seems i'll have to get busy on my end :wink: much respects RR you are the reason I bought over 2000ft. of poly tubing! Thanks again and GL everyone.


Edited by dcyans (07/18/04 10:32 AM)


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Invisiblesci33
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Re: clone from dried caps [Re: hyphae]
    #2902746 - 07/18/04 02:06 PM (19 years, 6 months ago)

I don't think this technique was meant as any type of long term storage, as you said, the distilled water tek is a good one for that purpose. I would think this thread was just meant for reviving a strain if the dried specimen was all that person had to work with. Risky, but proven possible. My bulk substrate is getting colonized right now.


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Re: clone from dried caps [Re: sci33]
    #2913046 - 07/21/04 04:53 PM (19 years, 6 months ago)

I can see that :wink:thanks anyway for the heads up? :wink: This was said for a friends ears. :wink:


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Re: clone from dried caps [Re: hyphae]
    #2947623 - 07/31/04 06:58 AM (19 years, 6 months ago)

Healthy looking, sporulating mushrooms from revived dried mycelium transferred to millet + rye, cased with verm/coir:


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Invisiblesci33
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Re: clone from dried caps [Re: Pinback]
    #2948861 - 07/31/04 02:00 PM (19 years, 6 months ago)

And an update on my progress:

The tissue from the agar drop was transfered to some peroxidated agar, on which it had a really strong resistance to the peroxide and ate it up quickly. From there went to jars of WBS, to a tray of worm castings / straw. It's finishing up the casing layer (very rhizomorphic) and will be fruited next week.


--------------------
Looking for:
Ps. weilii, Ps. samuiensis, Ps. hispanica, Pan. cyanescens (any strain I don't have)

Have:
Ps. cubensis, Ps. tampanensis, Pan. cyanescens (Australian, Tai-KS, Hawaiian, Jamaican, Ban Lipa Noi)

PM for a trade.


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Re: clone from dried caps [Re: Pinback]
    #2954098 - 08/01/04 10:35 PM (19 years, 6 months ago)

Quote:

Pinback said:
Healthy looking, sporulating mushrooms from revived dried mycelium transferred to millet + rye, cased with verm/coir:





Ok listen up guys like I said before I'm not saying dried myc won't produce. What I said is that I was referred to this thread by someone  as a means of long term storage, I have revived dried myc before that was dried for a short peroid of time. Cloning from a dried caps is not a question! :wink: GL and sorry for interrupting this fine thread. :laugh:


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Re: clone from dried caps [Re: vinsue]
    #2976099 - 08/07/04 05:11 PM (19 years, 6 months ago)

Glad you guys got the bugs worked out. This is a great way to store cultures.


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Re: clone from dried caps [Re: RogerRabbit]
    #2976312 - 08/07/04 06:22 PM (19 years, 6 months ago)

I wouldn't go that far... I still think that spores are germinating from the caps or that they are incompletely dried.

It wouldn't surprise me too much to find out that the mycelium can withstand drying with out dying though. It would be an excellent survival technique.

I highly doubt that a vacuum desicated or freeze-dried mushroom could be regenerated though. I also doubt that normaly dried mushrooms are viable for very long.


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Re: clone from dried caps [Re: fastfred]
    #2976502 - 08/07/04 07:46 PM (19 years, 6 months ago)

take a look at that cluster of shrooms they all look the same he cloned it.

pinback: did the pinset/flush look and perform like a clone??


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Re: clone from dried caps [Re: phucknut68]
    #2978449 - 08/08/04 02:08 PM (19 years, 6 months ago)

Well I was doing a petri yesterday and did one using a dried shroom thats at least 2 years old as thats when I bought it. Cracker dry and has remained that way in an airtight jar with silica gel. I cut a peice of the gill, a peice from inside the cap and a peice from the base of the stem, the soft spongy part and threw them all on a plate. I dont really care if its a spore or the shroom that goes if it goes, which I doubt completly, but took only a second to try. I just would like to be able to produce these again, not sure what they were but they were very large, and very potent. Well I'll be posting if it ever shows anything.


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Re: clone from dried caps [Re: vinsue]
    #2979280 - 08/08/04 08:24 PM (19 years, 6 months ago)

Totally, completely dried mushroom mycelia will grow again. It's a fact and not to be argued. It happens every year in the wild. Look at oyster mushrooms. In my area, there is NO rain from about mid May until early October. The forest fire danger is very high, and all the dead wood in the forest is cracker dry. If you cut into a log, as I did last week, you will see the mushroom myc, and it's as dry as a bone and looks dead. However, come October when the rains start, there will be oyster mushrooms by the truckload popping up from nearly every fallen tree, all over the forest. It happens every year. Culture banks store dried mycelia long term as a way of preserving isolated strains of edibles. The dried mycelia grows again when re-hydrated. Those aren't spores germinating, it's mycelium. Besides, I challenge anyone to grow oyster or shiitaki mushrooms from spores.


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Re: clone from dried caps [Re: phucknut68]
    #2979791 - 08/08/04 11:18 PM (19 years, 6 months ago)

Quote:

phucknut68 said:
take a look at that cluster of shrooms they all look the same he cloned it.

pinback: did the pinset/flush look and perform like a clone??




I don't think it's any unequivocal evidence either for or against:



The thing that makes me believe it's not spores is simply because it was an agar wedge I used. There is of course a (IMO small) possibility that cubensis spores in the air snuck into the petri dish where the wedges were drying, and landed on them. What speaks against this is that there are lots of other spores and bacteria in the air that could just as likely have landed on the wedges, but, there were no contaminations.

Nothing dried at room temperature will be "completely dry", there's always a small amount of water left, but I don't really get that argument. The wedge (dried for 55 days) was very hard, and actually split into two pieces when I speared it with a scalpell. I don't think it will get much drier than that at RT.

From what I gather, only four of the posters in this thread has actually tried this. I would be great if more did, especially the more skeptic ones.


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Re: clone from dried caps [Re: Pinback]
    #2980098 - 08/09/04 12:54 AM (19 years, 6 months ago)

A dried agar wedge?
How did you dry and store it ?

I wonder how long the dried mycelium would be viable ...
Would be a new method for long-term storage ...

Maybe one could make sure that no spores are germinating by using peroxidated agar ?


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Re: clone from dried caps [Re: ragadinks]
    #2980366 - 08/09/04 02:28 AM (19 years, 6 months ago)

I cut a wedge from a colonized plate, put it in a petri dish (the kind that doesn't make a perfect seal, I don't know the english word for it) and put it on a shelf away from direct sunlight, but still not dark. It was simply kept there for the entire time. I'm guessing the temperature was between 20 and 24?C.

It would really be interesting to see if it even survives on peroxidated agar. The reason vegetative mycelium survives, but not spores, is that the mycelium produces catalase, an enzyme that breaks down hydrogen peroxide. It is possible that the catalase might be denatured and no longer able to protect the cells when the mycelium is dried, but the only way of finding out is trying. Too bad it takes such a long time...


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Re: clone from dried caps [Re: Pinback]
    #2980375 - 08/09/04 02:31 AM (19 years, 6 months ago)

Hmmh, I guess the catalase won't survive the drying process, but as you mentioned: You only can be sure when you try it out.


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Re: clone from dried caps [Re: ragadinks]
    #2980606 - 08/09/04 05:56 AM (19 years, 6 months ago)

>Totally, completely dried mushroom mycelia will grow again. It's a fact and not to be argued.

I remain skeptical...

>It happens every year in the wild.

Do you have a citation for that? Perhaps it happens in the desert with certain species, but I've never read in any life cycle description that it actually dries out completely.

Culture banks store dried mycelia long term[...]

I don't mean to be argumentative, but could you provide some more information on that? Culture banks usually freeze samples in liquid nitrogen or keep them on slants. I've heard of them freeze-drying specimins, but not with the hope of reanimating them.

Could somebody describe the media they are using to get these results? The only mention I see is PDY... I would suggest that this is not the media to use as the yeast could contribute to the reanimation effect.

A foaf tried a similar experiment with LB (admittedly a shitty media) and did not get any growth.

-FF


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Re: clone from dried caps [Re: fastfred]
    #2980783 - 08/09/04 08:10 AM (19 years, 6 months ago)

In an earlier post in this thread, there is a link to a document from a culture bank describing this. The title of the document is "Instructions for Reviving Freeze-Dried Fungal Cultures"! Sorry, I don't have any specific details about the process, but it shows that it is actually done by professionals.


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Re: clone from dried caps [Re: Pinback]
    #2980914 - 08/09/04 09:28 AM (19 years, 6 months ago)



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Re: clone from dried caps [Re: fastfred]
    #2981290 - 08/09/04 11:18 AM (19 years, 6 months ago)

The reason freeze drying works for long term is that metabolic activity is almost stopped. Eventually a specimen will die if something is not done to stop this activity and we in our limited knowledge as humans cannot revive anything that is dead (actually dead) :wink: We can preserve and revive without question. Dried specimens over 1-20yrs. has not to my knowledge been successful in reviving, I'm open to documentation although, as apparently even after 55 days a "dried" specimen still is active metabolically. Like I first said I do not see this as any type of long term solution although thats not what the thread started out as anyway. :wink: GL


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Re: clone from dried caps [Re: hyphae]
    #2992448 - 08/11/04 06:10 PM (19 years, 6 months ago)

Well I grew green. I could try to dip in H202 and try again but I have not the will nor patience for that so good luck to you all with it :smile:


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Re: clone from dried caps [Re: vinsue]
    #2999989 - 08/13/04 09:59 AM (19 years, 6 months ago)

Coming in late on this, but FWIW...

An aquafriend of mine re-animated some shake, from a 3+ year old bag, in honeywater.  This shake had been stored in a ziplock, with dessicant, at room temperature the entire time. 

Marginal, but definite mycelium growth. (more than likely spore germination & not clone, but still a bit exciting).  This particular jar of honeywater was forgotten about for over a year until he recently cleaned his fridge.  That same jar had a rather large culture in it, quite a bit larger than originally stored. 

Instead of tossing it, he decided to see if it was still viable.  He let the jar warm in a cabinet for ~12 hrs, innoculated some sterilized rye, and has mycelial growth in just 2 days  :smile:
Tough stuff.


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Re: clone from dried caps [Re: Aquaman]
    #4770204 - 10/07/05 05:25 PM (18 years, 4 months ago)

Sorry to bump this old thread, but I have some new data.

A little over a year ago, I took small samples of colonized grain and bird seed and put it in sterilized test tubes. I plugged the opening with sterilized cotton, and put the tubes on a shelf where they resided in ambient light at room temperature. Note that only a small amount was used to ensure fast drying.


Some two weeks ago, I removed a few kernels and put them on an agar plate (standard PDYA plus 0.024% H2O2). This is what the plate looked like yesterday:


It does seem a bit fishy - at least on the left side. It looks better today though, the far left mycelium looks very similar to the right side on the picture. I have transfered a few wedges of the good looking side to a jar with grain to test it for fruiting.

To sum it up, the mycelium has survived after having been dried for >380 days, and could be revived on peroxidated agar.


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Re: clone from dried caps [Re: Pinback]
    #4770623 - 10/07/05 07:55 PM (18 years, 4 months ago)

Thanks for the update and picture. I've long believed that dried mycelium will outlast even spores. You culture above looks like it was from multispore with two distinct substrains saved. That's the difference you're seeing.
RR


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Re: clone from dried caps [Re: Pinback]
    #4770751 - 10/07/05 08:37 PM (18 years, 4 months ago)

Quote:

Pinback said:
Sorry to bump this old thread, but I have some new data.





Hey, thank you for giving it that bump.
I was just asking RR if he could find it,lol.


Doc

:cool: :thumbup:


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Re: clone from dried caps [Re: RogerRabbit]
    #4772149 - 10/08/05 07:12 AM (18 years, 4 months ago)

You are correct, I was almost certain that it was a clone, but when I checked my notes it was from multispore. :blush:


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Re: clone from dried caps [Re: Pinback]
    #4776105 - 10/09/05 08:09 AM (18 years, 4 months ago)

totally amazing...

I thought this property was exclusive only to sclerota producing myceliums? Cool.

Learn something RADICAL today... lol


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Re: clone from dried caps [Re: Terantula]
    #4776478 - 10/09/05 10:22 AM (18 years, 4 months ago)

Great work! Could be a good way for people to trade, if they can legally do it.


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Re: clone from dried caps [Re: blackout]
    #4785395 - 10/10/05 10:57 PM (18 years, 4 months ago)

I can attest the fact that dried cubenisis mushroom can grow on agar, because I have personally done it.


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Re: clone from dried caps [Re: Pinback]
    #4800182 - 10/13/05 06:52 PM (18 years, 4 months ago)

Question is that first pic taken at the end of a years storage Pinback?
RR I truely believe nature had a reason for creating such a hard surface for those of spores, I myself don't believe dried tissue could even come anywhere close but I'd love to be proven wrong! :wink:


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Re: clone from dried caps [Re: hyphae]
    #4801713 - 10/14/05 12:01 AM (18 years, 4 months ago)

No, it was taken after approximately 1.5 months of storage. But since the test tube looked exactly the same I didn't bother taking a new photo.


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Re: clone from dried caps [Re: Pinback]
    #4802504 - 10/14/05 05:57 AM (18 years, 4 months ago)

Just curious as my stored myc (on agar) has looked quite depressed and very dried out after a year in storage. I think grain would work better as well as a properly sealed culture as I had a couple out of many that dried so bad the agar had dried hard as a rock and the myc virtually disappeared. Damn that is some excellent looking myc you got going there with absolutely no signs of decreased viability. Nice job!


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Re: clone from dried caps [Re: hyphae]
    #4802556 - 10/14/05 07:10 AM (18 years, 4 months ago)

I have a LC about 1 month old that is showing no growth yet, it was stored at room temp. If you were to inject a small amount of LC into a jar with a filter and dry it out with damprid, do you think it could be revived later.
Also is there any reason why an LC at room temp for a year should not last as long as grain stored for a year?


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Re: clone from dried caps [Re: blackout]
    #4803103 - 10/14/05 11:19 AM (18 years, 4 months ago)

I think the key here is slowing the metabolic activity as much as possible to extend nutes and yet keep the tissue culture in a sort of hiberation type state. Damn 1mo. and no signs of growth? I would think chances would be good a LC would provide cultures after a year as long as it is contam free.


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Re: clone from dried caps [Re: hyphae]
    #4835818 - 10/21/05 07:37 PM (18 years, 3 months ago)

try it. germinating the spores is more likely to happen though. i've accidentally frozen spore syringes for a month or more and they worked just fine when i thawed em out. my two cents.


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Re: clone from dried caps [Re: fungusflip]
    #5405607 - 03/15/06 08:17 PM (17 years, 11 months ago)

used peroxide for what?


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Re: clone from dried caps [Re: chillinmush04]
    #5405715 - 03/15/06 08:38 PM (17 years, 11 months ago)

You can make spore prints with a year old dried mushroom? Does this mean I can just go to my local dealer instead of ordering syringes over the net?


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Re: clone from dried caps [Re: jonnyjonjonjon]
    #5408794 - 03/16/06 02:55 PM (17 years, 10 months ago)

technically, you could take a dry piece from the inside of the dry mushroom and place it on agar. if you did it enough times, you'd get one that started growing again.

practically, it wouldn't be worth the effort.


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Re: clone from dried caps [Re: shirley knott]
    #5408877 - 03/16/06 03:14 PM (17 years, 10 months ago)

that's debateable, imho.
see myc. syringes from dried tissue

picture


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Re: clone from dried caps [Re: Hippie3]
    #5409138 - 03/16/06 04:18 PM (17 years, 10 months ago)

let's wait and see what RR says. i know you respect each other  :rolleyes:


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Re: clone from dried caps [Re: shirley knott]
    #5409553 - 03/16/06 05:53 PM (17 years, 10 months ago)

rodger already commented on it,
in that very thread i linked.
Quote:

11-07-05, 02:01 #21
Rodger
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Imagine that. Just a year ago, nearly everyone thought it's impossible to revive dry tissue. Nice stuff.
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Re: clone from dried caps [Re: Hippie3]
    #5410621 - 03/16/06 11:04 PM (17 years, 10 months ago)

Very fine work Pinback. Nice indeed!


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Re: clone from dried caps [Re: Hippie3]
    #5411673 - 03/17/06 08:04 AM (17 years, 10 months ago)

Quote:

Hippie3 said:
that's debateable, imho.
see myc. syringes from dried tissue

picture




What exactly is debatable? Mycologists and tissue banks have been storing dry tissue cultures long term for years. It's the best storage method. Lasts many times longer than spores or 'live' tissue.
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Re: clone from dried caps [Re: RogerRabbit]
    #5411694 - 03/17/06 08:10 AM (17 years, 10 months ago)

also see
http://forums.mycotopia.net/showthread.php?t=6152

what is debatable is her statement-
Quote:

it wouldn't be worth the effort.




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Re: clone from dried caps [Re: Hippie3]
    #5411919 - 03/17/06 09:14 AM (17 years, 10 months ago)

oh sorry, misunderstood. I recently opened up a vacume sealed bag of mushies that was three years old. They started growing again on agar with zero contaminants. I'm thinking the dry tissue outlived the contaminant spores that were certainly on them. They were air dried for a few days before being put on desiccant and sealed, so it's a bet they were exposed to contamination. The dry fruits I've done this with that were one year old, seemed to grow a lot of trich and cladosporium, requiring several transfers to get a clean culture. Perhaps the key is to wait out the lifespan of the contaminant spores?
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Re: clone from dried caps [Re: RogerRabbit]
    #5412818 - 03/17/06 01:32 PM (17 years, 10 months ago)

I wonder if the vacuum sealing process had a lot to do with the contaminate spores not being able to hydrate and then germinate?


Edited by hotnutz (03/17/06 02:32 PM)


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Re: clone from dried caps [Re: RogerRabbit]
    #5473480 - 04/03/06 03:49 AM (17 years, 10 months ago)

Hi Roger
I am not aware of any professional mushroom farm or culture stockist who uses dried mycelium to store them. Not saying that it doesnt happen, but it isnt as common as your post seems to suggest.
I am aware of the processes used in a number in Australia and they all use - without exception - refridgerated slants and purchase without exception slants.

In terms of its viability though - most woodlovers produce heavily rhizomorphic mycelia which every year in teh summer months dries out completely only to reinvigorate when the rains and cold snaps come - subaeruginosa seems to do this particularly well and ive often wondered whether it is the spores or the dormant mycelia on the wood debris that results in the return of subs at a patch each year.
i wasnt aware that cubensis could do this, but one i thought about it it makes perfect sense.

If mushroom farms arent using this method and its perfectly viable maybe they should be :wink:


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Re: clone from dried caps [Re: Zen Peddler]
    #5488936 - 04/07/06 03:53 AM (17 years, 10 months ago)

I once bought dred shiitake mycelium on rye, but never was able to germinate it.

You can get it here
http://www.b-and-t-world-seeds.com/a1.asp?title=Fungus+Mycelium&list=184


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Re: clone from dried caps [Re: Anno]
    #5495061 - 04/08/06 11:26 PM (17 years, 10 months ago)

well im going to try and bring some old dried cloning material back to life - wish me luck


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Re: clone from dried caps [Re: snowspeed1]
    #5513024 - 04/13/06 05:12 PM (17 years, 10 months ago)

if i recall correctly from my biology class, cell walls do rupture when frozen,but not for fungi, as the cell walls for fungi are not enclosed and share all of the cell contents, cytoplasm, mitochondria, water ect. so the water has room to expand without casuing damage check out this pic of a mushroom cell. notice how it is open on two different ends.



Edited by redshadow (04/14/06 05:16 PM)


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Re: clone from dried caps [Re: redshadow]
    #7754604 - 12/13/07 04:46 PM (16 years, 2 months ago)

The stuff in this thread is really fascinating!  Necromancing this one because someone may find it interesting too :smile:


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Re: clone from dried caps [Re: Remanifest]
    #7754934 - 12/13/07 06:04 PM (16 years, 2 months ago)

you can do this for sure... thats how I store ALL my cultures... bone dry in a drawer. 

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6886717/an/0/page/0

theres the link to how I make my lids that allow me to dry out the cultures that are grown on grain.  if you go down the page, you'll see how I do it.  no one even replied to the thread. lol  theres a gold mine in that link and everyone seems to be just stepping over the entrance. hehehe..  the thing is (and you'll see this if you read that thread), after they are dried out and you retrieve the myc for use, you can just inject about 10 cc's of sterile water into the master jar that contains the dried out grain and myc and the grain will suck the water up and the myc will start growing again to be used once again... it's like you never even used the master jar.  best thing I've come up with.  :thumbup: ....so far.

fahtster


Edited by fahtster (12/13/07 06:10 PM)


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Re: clone from dried caps [Re: Remanifest]
    #7762445 - 12/15/07 04:40 PM (16 years, 2 months ago)

I almost forgot about this thread.I was a member here less than 2 weeks when I posted that...


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Re: clone from dried caps [Re: vinsue]
    #7768181 - 12/17/07 01:47 AM (16 years, 2 months ago)

Quote:

vinsue said:
I almost forgot about this thread.I was a member here less than 2 weeks when I posted that...




did the method work for you? :mushroom2:


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Re: clone from dried caps [Re: Floyd_]
    #7793434 - 12/23/07 06:25 PM (16 years, 1 month ago)

Believe me or not I grown mushrooms from dessicated one. If I have time I will post some picture and the procedure that I used.
It is no easy because the presence of bacteria (if not well preserved) but by continuous isolation you can grow healthy mushrooms, it takes time and care but it can be done.


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Re: clone from dried caps [Re: sven77]
    #7809698 - 12/29/07 03:19 AM (16 years, 1 month ago)

^ That sound pretty interesting. Please post some pics if you have the time.


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Re: clone from dried caps [Re: MayorMcCheese]
    #7810030 - 12/29/07 09:15 AM (16 years, 1 month ago)

I find it very easy myself. 9.9 out of 10 times there will be a contaminate present along with the mushroom culture. Most are simple to get away from with the exception of mucor. Mucor can be a real pain to get away from. You have to be on top of your transfers for sure. One slip in your transfers and you're back to square one when it comes to getting away from mucor.


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Re: clone from dried caps [Re: Hotnuts]
    #7850798 - 01/08/08 09:57 PM (16 years, 1 month ago)

I think someone needs to write up a tech about this type of process. I recently took a over 6 month old BRF cake that was just sitting in an old grow chamber, and brought the mycelium back to life in a LC style solution. I wish I would have known about this sooner.


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OfflineHotnuts
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Re: clone from dried caps [Re: 89ford]
    #7851216 - 01/08/08 11:13 PM (16 years, 1 month ago)

The process is exactly the same as cloning a live mushroom. No differences at all, with the exception that dry mushrooms that have been sitting around for some time may harbor more contaminates. That's about it really. Very easy for the most part.



Edited by Hotnuts (01/09/08 10:47 AM)


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OfflineSuchSmartMonkeys
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Re: clone from dried caps [Re: Joshua]
    #7860760 - 01/10/08 05:41 PM (16 years, 1 month ago)

Quote:

Joshua said:
RogerRabbit says he has accomplished this, I guess he might be the man. Although I did see a pic of him wearing a bra and bunny ears once :lol:

I will give it a try.

Joshua




that bra happened to be fruiting some pretty nice looking cube, if i do remember correctly.... I assumed RR had to have been ripped out of his head o be dancing around like that....how could you not be?


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Re: clone from dried caps [Re: Pinback]
    #8570960 - 06/27/08 12:54 PM (15 years, 7 months ago)

A late update again; I took a grain from the batch described in this post and placed it on PDYA. It had been stored dark, but at ambient temperature. No growth whatsoever one week later.

I also had a mycelial suspension (mycelium scraped from agar and suspended in water) from 2001-2002 (not sure exactly since the label had come off!) that I placed on agar. No growth whatsoever one week later.

I'll update later if growth is resumed, but I don't count on it.


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Re: clone from dried caps [Re: Pinback]
    #8571149 - 06/27/08 01:45 PM (15 years, 7 months ago)

It can easily take a week or two to get started again.
RR


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Re: clone from dried caps [Re: vinsue]
    #9537230 - 01/03/09 10:54 AM (15 years, 1 month ago)

Does anyone think i could just go and buy a eighth of mushrooms, put a cap in agar or rye or vermiculate and wind up growing some nice healthy shrooms? "cap may be dryed"


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Re: clone from dried caps [Re: 4Sub2Noize0]
    #9542082 - 01/04/09 04:45 AM (15 years, 1 month ago)

Not in vermiculite (which doesn't contain nutrients, it's just used to add water reservoirs to substrates and casing layers) and not by dropping it onto rye. This technique requires lab skills. You'll want to use agar in a petri dish, because an 1/8 of dried shrooms will be full of contaminants. So what you want to do is to get P. cubensis mycelium to start growing again, not bacteria or mold or whatever. It's more than likely that you'll have various things germinating and growing side by side in the first petri dish, so you want to try and isolate clean mycelium and transfer it onto another petri dish. Repeat if necessary until all you see is healthy mycelial growth, nothing else. THEN you can cut out wedges and transfer them to your substrate or an LC jar.
If you think you are up to the task and can cater for a sterile environment then, yes, it is possible to do just what you described.
Growing mushrooms is different from growing plants. You cannot just "stick a seed in the ground". Then again, mycelium and spores are much sturdier than most plant seeds are and thus such a wicked trick will work. Provided you have the skills set.


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Re: clone from dried caps [Re: vinsue]
    #9542725 - 01/04/09 09:16 AM (15 years, 1 month ago)

This thread is still going ?? I was a member less than two weeks when I posted it . . .  P.S.  Never did do the the agar experiment:shrug: . . . :peace:


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Re: clone from dried caps [Re: vinsue]
    #9543037 - 01/04/09 10:36 AM (15 years, 1 month ago)

Quote:

Not in vermiculite (which doesn't contain nutrients, it's just used to add water reservoirs to substrates and casing layers)




Don't be so sure about that.  Mushroom mycelium will fully colonize vermiculite and fruit from it.  It's a bulk substrate material for many species, even though it's mineral.  I've seen brf cakes which are 2/3 verm shrink down to the size of dollar pancakes after 7 or 8 flushes.  Obviously, the mycelium has consumed the verm too.
RR


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Re: clone from dried caps [Re: RogerRabbit]
    #9543087 - 01/04/09 10:49 AM (15 years, 1 month ago)

You know, I wouldn't be surprised if this would be the case. Mixed with other minerals it is used as a veterinary feeding additive:



I do believe that it contains minerals that are beneficial, but I don't believe that it would be so ideal as a bulk substrate all by itself. And certainly not for reviving some dry shrooms obtained on the streets from it.


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Re: clone from dried caps [Re: German Kahuna]
    #9550092 - 01/05/09 09:26 AM (15 years, 1 month ago)

Vermiculite is not completely inert. Perlite however is.


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Re: clone from dried caps [Re: HerbBaker]
    #9553825 - 01/05/09 08:30 PM (15 years, 1 month ago)

I currently have 12-15 pieces of 2 year old dried caps and stems in PC'ed distilled water.  The pieces were triple washed with PC'ed distilled water as per RR's "tek" found on several post.  It has been 2 weeks now and the water has turned light brownish in color and filled with small floating blobs similar to past lc's that i've made.  Several pieces have started to get a little fluffy, these will be transfered to agar. I'll try to get some pics.  What do you all think the floaters in the water may be?  Contaminants, mycelium from spores or the pieces?


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Re: clone from dried caps [Re: PennyPincher]
    #9557251 - 01/06/09 11:35 AM (15 years, 1 month ago)

Quote:

What do you all think the floaters in the water may be? 



There's no way to tell.

Transfer the pieces with mycelium to agar, and be prepared to make a series of transfers to get live mycelium away from any contaminants.  Since it's hard to tell contaminant fungi from mushroom mycelium, transfer each growth you see to new plates.  That way, some will end up being contamination, but others will hopefully be mushroom mycelium.
RR


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Re: clone from dried caps [Re: RogerRabbit]
    #9558429 - 01/06/09 03:56 PM (15 years, 1 month ago)

RR, is this stuff floating in the water common when you use this method?


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Re: clone from dried caps [Re: PennyPincher]
    #9560985 - 01/06/09 10:15 PM (15 years, 1 month ago)

I've seen mushroom tissue that floats and some that sinks.  If that's not what it is, then I don't know.
RR


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Re: clone from dried caps [Re: RogerRabbit]
    #9594221 - 01/12/09 11:00 AM (15 years, 1 month ago)

hi all ive been reading this thread for some insight. I live in cali so I cant order any spores so I went ahead and bought some agar and petri dishes at the local teachers supply store. (ripoff) but anyways a buddy of mine gave me a dried mushroom of what looks like a pe strain. whell anyways I made up my petri dishes, and cut out some samples and but them on the dish. its been about 3 or 4 days and this is what I got. my question is "when do I cut out growth and transfer it onto another dish? hou long should I gro it out". thanks for your input


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Re: clone from dried caps [Re: califarmer619]
    #9594604 - 01/12/09 12:43 PM (15 years, 1 month ago)

Sorry to say it, what you have there are bacteria and molds.


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Re: clone from dried caps [Re: califarmer619]
    #9594619 - 01/12/09 12:46 PM (15 years, 1 month ago)

It is a bit hard to tell from your photo, but the dark specks of mushroom tissue with just a bit of fuzz around them are likely the good stuff and look right considering the age.  You should transfer away from the large slimy blobs (bacteria) immediately for best results.  You may have to do this more than once before getting a clean culture.  Good luck!


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Re: clone from dried caps [Re: califarmer619]
    #9595207 - 01/12/09 02:18 PM (15 years, 1 month ago)

In case your confused with what Workman just told you. The two upper right pieces, big w/ small piece next to it. I would transfer the mycelium around the little piece. That would be a good start.  After you transfer the mycelium place a sterile 3/4 inch by 3/4 inch piece of cardboard right next to the wedge and let the mycelium grow up above it then grab that mycelium with your loop.( this doesn't always work but most of the time it does) It will help separate it from bacteria if there is any present. I do this all the time with cultures that have bacteria in them and it works for me very well. Was there no gill tissue on the specimen you found? You can always just place a piece of gill tissue on the agar and try to get the spores on it to germinate.

Lipa


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Re: clone from dried caps [Re: lipa]
    #9595545 - 01/12/09 03:20 PM (15 years, 1 month ago)

yea I had been gifted only one complete mushroom with the veil partially intact. does anyone know of a thread that has good picture of agar cultures or young ones for reference. thanks for your help


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Re: clone from dried caps [Re: califarmer619]
    #9609151 - 01/14/09 03:44 PM (15 years, 1 month ago)

heres a shot of amother dish


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Re: clone from dried caps [Re: califarmer619]
    #9612985 - 01/15/09 05:51 AM (15 years, 1 month ago)

That looks excellent.


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Re: clone from dried caps [Re: Pinback]
    #24562733 - 08/18/17 12:36 PM (6 years, 5 months ago)

Hi!

Time to bump up this thread! I've just bought some dried mushrooms for the second time online and would just like someone to confirm with me if I can use any of these mushrooms/parts and just put on a petri dish with agar and with some luck it will start to produce mycelium which I then later can plant in a substrate?:ooo:

[flash=,][/flash]


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Re: clone from dried caps [Re: Zelikoo]
    #24563174 - 08/18/17 03:36 PM (6 years, 5 months ago)

You can make your own new thread too. But starting from dried is usually an uphill battle even for seasoned cultivators.


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Re: clone from dried caps [Re: bodhisatta]
    #24567867 - 08/20/17 04:50 PM (6 years, 5 months ago)

13+ years ago.  :wowz:

This was the first thread I ever made.
silly noob :burke:

I never did clone those caps, just ate them and
went ahead and ordered some spore syringes.

:mushroomgrow::sporedrop: . . . :peace:


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