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Offlinejasonmobile
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Registered: 06/24/11
Posts: 9
Loc: mobile, alabama
Last seen: 13 years, 4 months
cloning PE with no results
    #14723687 - 07/06/11 12:47 AM (13 years, 6 months ago)

I recently tried unsuccessfully to clone a Penis Envy fruitbody. Instead of using agar, I transferred inner tissue directly to 3 sterilized quart jars of organic rye berries (using glove box).  I used the standard water/rye ratio floating around out there.
        No results.  It's day 8 now and still no sign of mycelium, so I've given up hope on that project;  however, I'd like to learn from this mistake, but I can't figure out what I did wrong...
        At first, I thought my tissue samples were just too small, but after looking at similar agar teks I realize those tissue samples are small also.
        The only thing that I know I did wrong involved the jar lids during pressure cooking.  I forgot to invert the lids with rubber seal facing up (they were facing down instead), so the lids were kinda stuck on there with considerable vacuum.  I don't know if this maybe caused the substrate to become over dried...I am only speculating here...
        Any thoughts on what I could have done wrong?

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OfflineSTR80180
Stranger

Registered: 12/13/10
Posts: 142
Last seen: 4 years, 7 months
Re: cloning PE with no results [Re: jasonmobile]
    #14723902 - 07/06/11 01:57 AM (13 years, 6 months ago)

i doubt throwing a piece of fruitbody into a jar or rye will take off because agar is more nutrient than your rye berries, but it still might 8 days isnt that long, you probably would have seen growth around the tissue from the outside by now though, You could make a clone LC by placing a some inner fruitbody tissue into some sterilized honey water in a glovebox under sterile conditions and then use that as your inocculant (once you have seen signs of growth) for future grows, if it doesnt contaminate, it will last a few months in the fridge although it is highly susceptible to contaminants and its hard to tell if it is contaminated until your jars all turn green, but if you get a clean one you can make endless new jars with tiny amounts of your LC

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Invisiblewildernessjunkie
Reshitivest
I'm a teapot


Registered: 06/13/10
Posts: 8,118
Loc: HTTP 404 Not Found
Trusted Cultivator
Re: cloning PE with no results [Re: STR80180]
    #14724088 - 07/06/11 03:35 AM (13 years, 6 months ago)

Quote:

STR80180 said:
agar is more nutrient than your rye berries





Im going to disagree with that. Ive had agar pin and grow before. From everything Ive seen and experienced, this always results in a single small shroom. Agar is great for isolating, and culturing mycelium, but for growing an actual fruit body its not so good.

I have always thought about doing exactly what you have done, though I never have. So Im curious as to how this turns out. 8 Days is very little time, for an excised sample to begin growing. Give it another week, and see what happens. Maybe an "Old Hand" can chime in and give you some guidance, or school me on how to STFU.

I hope it works out for you.

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Invisibleanonjon
Partially Right

Registered: 11/03/08
Posts: 6,322
Re: cloning PE with no results [Re: wildernessjunkie]
    #14724296 - 07/06/11 05:46 AM (13 years, 6 months ago)

Problem was your rye prep. It needs to be pretty moist for this to work.

Here's a tek on the procedure.
http://myco-tek.org/showthread.php?94-Cloning-to-Grain-with-Open-Source

Some pics:



BAM!


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

Edited by anonjon (07/06/11 05:48 AM)

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Offlinek00laid
NEMO
 User Gallery

Registered: 05/03/10
Posts: 19,636
Last seen: 1 year, 5 months
Re: cloning PE with no results [Re: anonjon]
    #14724592 - 07/06/11 08:10 AM (13 years, 6 months ago)

it wouldnt let me choose you as my referrer, jon


--------------------
AMU - AMU Q & A - MyVideo Teks!

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Offlinejasonmobile
Stranger
Male


Registered: 06/24/11
Posts: 9
Loc: mobile, alabama
Last seen: 13 years, 4 months
Re: cloning PE with no results [Re: k00laid]
    #14725100 - 07/06/11 10:52 AM (13 years, 6 months ago)

thanks for the great advice anonjon.

it gives me reason to keep pushing ahead.

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OfflineFungi4TW
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Registered: 06/27/11
Posts: 40
Loc: Northwest Georgia
Last seen: 12 years, 4 months
Re: cloning PE with no results [Re: jasonmobile]
    #14728692 - 07/06/11 09:58 PM (13 years, 6 months ago)

I'll tell you how I have always cloned, and had great success.

I have a small blender that holds like 16oz. I put about 8oz of water in it, put the lid on, cover it with foil, and drop it in a big pot of boiling water. I let it boil for about 30 minutes, take it out and let it cool.

After it cools I put it in my glovebox, along with a scalpel, and a big ass mushroom with a fat stem (P.E.). The mushroom has been dunked in H202, and is wrapped in a H2O2 soaked towel. I cut away the outside layer from the stem, and take a big chunk from the center, and drop it in the blender cup.

Then I take it out of the glovebox, put it on the blender base and chop the stem piece up to very fine particles. I open the lid just enough to stick the needle from a 60cc syringe in, and suck up the clone water. You can use this to inoculate a bunch of jars.

I have had great success doing this. It's relatively quick and easy, too.


--------------------
Yes ma'am, that IS a screwdriver in my pocket!

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Offlineafrosheen
9Lives the cat
Male User Gallery


Registered: 03/06/10
Posts: 1,878
Last seen: 3 years, 3 months
Re: cloning PE with no results [Re: Fungi4TW]
    #14728837 - 07/06/11 10:27 PM (13 years, 6 months ago)

I've seen direct tissue transfers to grains work fine more than once. I wouldn't recommend it to everyone but it does work, with PE in fact.


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