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 BlueBruiser
Psilocin Technician
Registered: 03/14/02
Posts: 225
Loc: Mushroom planet
Last seen: 4 years, 1 month
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 Pan Cyan Cloning Technique from Fruit w/ pics 
#6209341 - 10/25/06 01:44 AM (17 years, 5 months ago) |
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So I was stoked about my first Pan grow found here. I wanted to take a clone and work with it but Pan mushrooms are so tiny I was wondering how I was going to take a clean tissue culture. I first tried the stem. I cut a piece of stem off, dipped it in alcohol (70%) and then in H2O2 and then into a liquid culture. All of them contaminated. Since I had tons of mushrooms I practiced surgical procedures in order to try and take some of the interior of the stem but was not able to do so easily because its very fiberous and tough. Then I tried the cap. I found the tissue there to be non-fiberous and very easy to scrape off. I did the following procedure and I successfully took 3 clean clones from the tiny Pan Cyan mushroom. Check it out and I hope it helps you guys.
Step #1
Using sterile technique take a scalpel and cut a thin slice of the SKIN of the cap. Just the skin, try not to go too deep since the cap is very thin in a Pan mushroom, to expose the soft interior tissue.
See how thin it is.
Step #2
Take a sharp pointed scalpel or similar object, again using sterile technique, and scrape some of the exposed white tissue from the middle of your slice. It should scrape off very easily. Like so.
Step #3
You now have a clean tiny piece of mushroom tissue. You can now transfer it to an agar plate or into a liquid culture. I simply touch the piece of tissue against a dry part of the wall of the liquid culture container carefull not to touch any droplets or the solution and it should stick. Close the container and give it a swirl and your clone should wash up into the solution. Incubate, inoculate, fruit and repeat. 
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vinzy_a
Stranger
Registered: 08/31/06
Posts: 201
Last seen: 14 years, 11 months
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Re: Pan Cyan Cloning Technique from Fruit w/ pics [Re: BlueBruiser]
#6209363 - 10/25/06 01:57 AM (17 years, 5 months ago) |
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so the tissue you got is reaaaaaaaaaaaalyyyy tiny? damn that must be hard..
try it on some h202 agar to fight off some contams or the cardboard tek..
cool cloning! 
oh btw, your grow was awesome!
Edited by vinzy_a (10/25/06 01:57 AM)
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RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 42,214
Loc: Seattle
Last seen: 1 year, 1 month
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Re: Pan Cyan Cloning Technique from Fruit w/ pics [Re: vinzy_a]
#6209657 - 10/25/06 06:08 AM (17 years, 5 months ago) |
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You can do the same thing with the stem. Just rip the stem length wise to split it in half, then gently scrape to get a tiny piece of mycelium. Never use alcohol on mycelium.
RR
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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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 memes
Blessed
Registered: 01/11/05
Posts: 27,785
Loc: In a Tree
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Re: Pan Cyan Cloning Technique from Fruit w/ pics [Re: RogerRabbit]
#6209672 - 10/25/06 06:16 AM (17 years, 5 months ago) |
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very nice, and beautiful grow too.
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Blutjager
Inhuman
Registered: 06/11/06
Posts: 9,220
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Re: Pan Cyan Cloning Technique from Fruit w/ pics [Re: memes]
#6209751 - 10/25/06 07:06 AM (17 years, 5 months ago) |
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I look forward To doing a pan grow in the future.Its always good to see information about them
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BlueBruiser
Psilocin Technician
Registered: 03/14/02
Posts: 225
Loc: Mushroom planet
Last seen: 4 years, 1 month
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Re: Pan Cyan Cloning Technique from Fruit w/ pics [Re: RogerRabbit]
#6211148 - 10/25/06 04:17 PM (17 years, 5 months ago) |
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Im sure you can, but I dunno if you have tried it on a Pan before. First off Pans are tiny which makes it a micromanagment thing and the stem is fiberous so when you scrape the inside of the pan the fiberous tissue tends to split along the fibers rather than easily scooping out a piece of tissue. The stem is difficult to work with because of the mentioned properties. The cap scrapes off tissue like butter, the stem like beef jerkey. I just wanted to show an extremely simple and successful way of cloning pans with out too much of a hassle.
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indospore
Stranger
Registered: 12/13/18
Posts: 2
Last seen: 4 years, 5 months
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 please help me [Re: vinzy_a]
#25684909 - 12/17/18 10:12 AM (5 years, 3 months ago) |
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Somebody please help me
How to make tissue from auricularia judae(Jelly Ear) muhroom to agar?
Please....
Edited by indospore (12/17/18 10:21 AM)
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