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Offlineblackout
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Dirty cloning method with H2O2, seems to have worked.
    #4368638 - 07/04/05 05:35 AM (11 years, 5 months ago)

I grew out some tiny shrooms. They dropped spores all over themselves. The stems were very thin so it would be difficult to open them up and get tissue that I was sure had no spores on it. I have no glove box or hood.

So I got 1/2 a stem and broke it up with my unwashed hands in open dirty air. I popped it all in a small glass jar with some broken glass and pure 3% H2O2. I shook it up and left for about 10mins. My idea was the spores would be killed and I could make a myc. tissue syringe. I sucked up 5cc of solution in a clean syringe through a self healing injection site and injected 2 jars.

2 days on, one of the jars shows growth in about 10 places, the other has what may be growth in 2.

If doing it again I would cut the stem up really fine with a razor or scissors.

I thought the strong peroxide may kill it. Does anybody know how long the tissue can survive in 3% peroxide? I still have some in it and will try again in a few weeks.


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OfflinekronnyQ
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Re: Dirty cloning method with H2O2, seems to have worked. [Re: blackout]
    #4368641 - 07/04/05 05:39 AM (11 years, 5 months ago)

H2O2 won't just destroy contams like a miracle cure, it only helps prevent contamination by a small percentage. I don't even use it anymore.

If your jars don't contam you are incredibly lucky or just really clean.


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Re: Dirty cloning method with H2O2, seems to have worked. [Re: kronnyQ]
    #4368676 - 07/04/05 06:28 AM (11 years, 5 months ago)

I know its not a miracle cure, but it is meant to kill all spores. I have heard of people working with peroxidated agar in open air with success. Its a very easy option if it does work. I put some in an LC but it shows no signs of growth yet. I thought the LC would have started quicker.


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Offlinepsilocyben
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Re: Dirty cloning method with H2O2, seems to have worked. [Re: blackout]
    #4369031 - 07/04/05 11:36 AM (11 years, 5 months ago)

i was reading that some had found that the initial dunk only proved as counterproductive to the growth of the culture as long as you were transferring to peroxidated media anyway.

i know for a fact that tissue will survive for weeks in peroxide.

it will take much longer to become acclimated to the medium, though.
:zoom:


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Re: Dirty cloning method with H2O2, seems to have worked. [Re: psilocyben]
    #4369097 - 07/04/05 12:21 PM (11 years, 5 months ago)

I thought the LC would take off quicker since I injected only about 2cc in a jar of about 500ml 2% malt extract. I figured it would dilute the H2O2 so its effects would be minimal.
How strong was the peroxide in which your tissue lasted weeks? I think I should leave it in the sunlight to degrade faster to H2O


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Offlinehyphae
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Re: Dirty cloning method with H2O2, seems to have worked. [Re: blackout]
    #4369156 - 07/04/05 12:51 PM (11 years, 5 months ago)

Ya weeks would be a misleading statement since the peroxide would have been broken down in a matter of days after introducing foreign matter. Whenever I have done tissue culture transfers I have always done a 20 min. peroxide (3%) dunk. It's great insurance IME as well as many others I know that use the same procedure. GL


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Amazon Shop for: ½ Pint Jars, Agar, Malt Extract

Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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