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OfflineDelay
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Registered: 10/29/13
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Tissue-specific cloning
    #19217094 - 12/02/13 12:38 PM (10 years, 1 month ago)

Hi,

I'm sorry if this question has been previously answered. I've been cloning mushrooms from the grocery store. I've been doing so by purchasing mushrooms who's stems have a piece of mycelium seemingly from the casing/substrate still attached, and excising the tissue from that portion of the produce. I figured this mycelium would work best, but can you clone mycelium from the upper portion of the stem or cap with equal results? Are those tissue as/more/less vigorous when cloned in comparison to one another?

Cheers,

Delay


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Offlinekrypto2000
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Re: Tissue-specific cloning [Re: Delay]
    #19217119 - 12/02/13 12:43 PM (10 years, 1 month ago)

You can clone from any part of a mushroom. The best thing to do is to cut it in have and take the part at the top of the stem right where it meets the cap.


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InvisibleFrankHorrigan
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Re: Tissue-specific cloning [Re: krypto2000]
    #19217128 - 12/02/13 12:45 PM (10 years, 1 month ago)

Quote:

krypto2000 said:
You can clone from any part of a mushroom. The best thing to do is to cut it in have and take the part at the top base of the stem




Fixed it for ya :wink:

OP, here is how to clone things with great success.


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

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


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AMU- Get an answer here -AMU


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Offlinekrypto2000
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Re: Tissue-specific cloning [Re: FrankHorrigan]
    #19217488 - 12/02/13 02:16 PM (10 years, 1 month ago)

Oops, thanks frank.


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Invisible36fuckin5
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Loc: Diving into Mystical Territori...
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Re: Tissue-specific cloning [Re: krypto2000]
    #19217527 - 12/02/13 02:24 PM (10 years, 1 month ago)

I personally like to clone from the cap. The flesh is easier to cut up.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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InvisibleFrankHorrigan
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Re: Tissue-specific cloning [Re: 36fuckin5]
    #19217531 - 12/02/13 02:25 PM (10 years, 1 month ago)

Cap works too.

I love the base of the stem though, I get the fewest number of contams that way.


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

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


Frank's tips and tricks. Updated on 3/21/14
AMU- Get an answer here -AMU


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OfflineDelay
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Re: Tissue-specific cloning [Re: FrankHorrigan]
    #19218384 - 12/02/13 05:38 PM (10 years, 1 month ago)

Quote:

FrankHorrigan said:
Cap works too.

I love the base of the stem though, I get the fewest number of contams that way.




Why would the stems be less likely to become contaminated???


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OfflinePsilicon
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Re: Tissue-specific cloning [Re: Delay]
    #19218510 - 12/02/13 06:03 PM (10 years, 1 month ago)

My guess would be that it's solid down there, leaving no room for bacteria to grow.


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InvisibleFrankHorrigan
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Re: Tissue-specific cloning [Re: Delay]
    #19220026 - 12/02/13 11:02 PM (10 years, 1 month ago)

Quote:

Delay said:
Why would the stems be less likely to become contaminated???




I didn't say they were less likely to become contaminated in general, just that I see the fewest contams when I take cuts from the center of the base of the stem.

Why? I don't know.


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

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


Frank's tips and tricks. Updated on 3/21/14
AMU- Get an answer here -AMU


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OfflineRogerRabbitM
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Re: Tissue-specific cloning [Re: Delay]
    #19220710 - 12/03/13 05:14 AM (10 years, 1 month ago)

You can clone from just about any fleshy part of the mushroom.  However, you're going to get less than ideal performance from grocery store mushrooms.  First, they've already expanded the mycelium, and then fruited it.  If you clone and start over, you're starting from old cell lines.  You'll be miles ahead to obtain a low-time culture.

When you clone, tear the stem or cap in half and carefully cut a tiny piece of mycelium from the inside tissue which has never been exposed to air.
RR


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OfflineDelay
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Re: Tissue-specific cloning [Re: RogerRabbit]
    #19221233 - 12/03/13 09:29 AM (10 years, 1 month ago)

Quote:

RogerRabbit said:
You can clone from just about any fleshy part of the mushroom.  However, you're going to get less than ideal performance from grocery store mushrooms.  First, they've already expanded the mycelium, and then fruited it.  If you clone and start over, you're starting from old cell lines.  You'll be miles ahead to obtain a low-time culture.

When you clone, tear the stem or cap in half and carefully cut a tiny piece of mycelium from the inside tissue which has never been exposed to air.
RR




Thanks, I'm simply cloning these grocery store mushrooms because I am a student and I am too poor to purchase cultures/spores, but I appreciate your input regarding senescence.

I was aware of the technique for cloning. I have a laboratory background, and I'm familiar with sterile procedures, which I employed as much as was possible. That being said, I don't have access to a flow/laminar hood, so I cloned within a circle of candles like an old school microbiologist. Furthermore, I entirely skipped agar, cloning straight to grain. It's been >10 days, clones are growing, and there are no visible contaminants as of yet. This was mostly an experiment. I cloned Lentinola edodes, Pleurotus ostreatus and Agaricus bisporus. I know it ain't over til the fat lady sings, so I'm still waiting for full colonization to gauge the success of my method.


Edited by Delay (12/03/13 10:30 AM)


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