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InvisibleLucid_Euphoria
Sojourner


Registered: 03/23/05
Posts: 1,045
Loc: Flip Side
Live Tissue to LC procedures?
    #10618057 - 07/03/09 09:27 PM (15 years, 6 months ago)

Ok I'm looking for proper procedures to to transfer live tissue from an outdoor patch shroom to an LC.

Please critique what I've got so far:

1. Pick the given specimen, place into plastic baggy
2. Place all materials in glove box
3. Let glove box settle
4. Flame Sterilize scalpel
5. Wipe outside of shroom with alcohol swab
6. Split shroom in half
7. Cut out a small chunk from the center, up near the cap
8. Dunk chunk into 10% bleach solution
9. Open LC jar & drop chunk in
10. Close LC

Anything you see wrong or that would increase my chances of success?


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PAN CYAN & AZURE FOR TRADE

"If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha

"Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-

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OfflineMushroomMaster
Horticulturalist
Male User Gallery


Registered: 07/02/09
Posts: 59
Loc: Deep Sea Lair
Last seen: 15 years, 6 months
Re: Live Tissue to LC procedures? [Re: Lucid_Euphoria]
    #10618087 - 07/03/09 09:38 PM (15 years, 6 months ago)

If its wild I don't know if that would work. That's like adding a contamination straight into the LC.

Your best bet in reproducing a wild specimen would be to drop a sample into agar and isolate or making a spore print. You can drop the tissue directly into a grain jar but it being wild means its covered in a fruit basket of problems.


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:mushroom2:PES Hawaiian  :mushroom2:Chit-wan  :thumbup:Grow Your Own Rye


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InvisibleLucid_Euphoria
Sojourner


Registered: 03/23/05
Posts: 1,045
Loc: Flip Side
Re: Live Tissue to LC procedures? [Re: Lucid_Euphoria]
    #10618118 - 07/03/09 09:45 PM (15 years, 6 months ago)

Quote:

it being wild means its covered in a fruit basket of problems.




Covered being the keyword there. A sample from inside the mushroom should be clean.

I have spores & could do an MS inoculation to the LC, but I wanted to try a "cloning" of a viable strain.


--------------------
PAN CYAN & AZURE FOR TRADE

"If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha

"Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-

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OfflineMushroomMaster
Horticulturalist
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Registered: 07/02/09
Posts: 59
Loc: Deep Sea Lair
Last seen: 15 years, 6 months
Re: Live Tissue to LC procedures? [Re: Lucid_Euphoria]
    #10618142 - 07/03/09 09:51 PM (15 years, 6 months ago)

If I were you I would Spore to LC and after you get an indoor sample I would try cloning to LC with that. Just because you get tissue from inside doesn't make it clean.

To be honest tissue to LC sounds like it won't work. Only reason I think that is because if the tissue breaks down you will have a small bit of rot floating around. That's just what I hypothesize.


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:mushroom2:PES Hawaiian  :mushroom2:Chit-wan  :thumbup:Grow Your Own Rye


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OfflineMushroomMaster
Horticulturalist
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Registered: 07/02/09
Posts: 59
Loc: Deep Sea Lair
Last seen: 15 years, 6 months
Re: Live Tissue to LC procedures? [Re: MushroomMaster]
    #10618153 - 07/03/09 09:53 PM (15 years, 6 months ago)

Also to truly clone or "isolate desirable characteristics" You should use agar. Then you can Isolate the part of the mushroom that made you want to clone it in the first place.

:shoosh:


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:mushroom2:PES Hawaiian  :mushroom2:Chit-wan  :thumbup:Grow Your Own Rye


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InvisibleLucid_Euphoria
Sojourner


Registered: 03/23/05
Posts: 1,045
Loc: Flip Side
Re: Live Tissue to LC procedures? [Re: Lucid_Euphoria]
    #10618214 - 07/03/09 10:11 PM (15 years, 6 months ago)

I'm aware of the differences between "cloning" and isolation.

Quote:

To be honest tissue to LC sounds like it won't work.




It does work. It's been done many times.


--------------------
PAN CYAN & AZURE FOR TRADE

"If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha

"Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-

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OfflineMushroomMaster
Horticulturalist
Male User Gallery


Registered: 07/02/09
Posts: 59
Loc: Deep Sea Lair
Last seen: 15 years, 6 months
Re: Live Tissue to LC procedures? [Re: Lucid_Euphoria]
    #10618223 - 07/03/09 10:14 PM (15 years, 6 months ago)

Well I wish you luck, I also use LC I think I will give it a shot.


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:mushroom2:PES Hawaiian  :mushroom2:Chit-wan  :thumbup:Grow Your Own Rye


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InvisibleCH HELL
Brain Sturgeon
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Registered: 10/02/08
Posts: 6,610
Loc: mars Flag
Trusted Cultivator
Re: Live Tissue to LC procedures? [Re: MushroomMaster]
    #10618322 - 07/03/09 10:50 PM (15 years, 6 months ago)

Sounds like you are making it to hard.  yes wipe the shroom down before it goes in the GB.  Have two LC's ready,  tear the shroom in half the center is sterile.  Take a peice from the base of the shroom(the white meaty part) drop it in an LC.  Then take a sample from the top near the cap again the white meaty part and drop it in an LC.
GL,
CH

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OfflineNineInchNails
Stranger

Registered: 03/01/09
Posts: 1,190
Last seen: 2 years, 12 days
Re: Live Tissue to LC procedures? [Re: CH HELL]
    #10618349 - 07/03/09 10:59 PM (15 years, 6 months ago)

If not straight to agar then a print to LC sounds like the best way to go.

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Offlinehyphae
born to grow
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Registered: 12/13/02
Posts: 6,228
Loc: the rain forests
Last seen: 13 years, 8 months
Trusted Cultivator
Re: Live Tissue to LC procedures? [Re: NineInchNails]
    #10621203 - 07/04/09 01:58 PM (15 years, 6 months ago)

I've done this many times personally the only difference is I use tissue from under the epidermal layer of the cap three or four small chunks. I place those chunks in a 3% peroxide bath for a few minutes then drop them in the LC.
FYI a tissue sample from a live fruit body is the best isolate you can have besides of course a better tissue sample from a better fruit body. LOL


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Getting the most out of your casings!, A pinning strategy.
Oyster Shell "Flour" $2 for 1lb. a hell of a deal :wink:
Not what is overlay but rather what overlay is
Gas Exchange vs. FAE

"We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"

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InvisibleLucid_Euphoria
Sojourner


Registered: 03/23/05
Posts: 1,045
Loc: Flip Side
Re: Live Tissue to LC procedures? [Re: hyphae]
    #10621437 - 07/04/09 03:03 PM (15 years, 6 months ago)

Quote:

a live fruit body is the best isolate you can have besides of course a better tissue sample from a better fruit body.




I thought fruit tissue could still sector? Meaning it isn't actually an isolate?

I could be wrong there just thought that's what I've read.

I did one for the hell of it to see if I'd succeed.


--------------------
PAN CYAN & AZURE FOR TRADE

"If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha

"Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-

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