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Invisiblegreys
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Registered: 07/16/06
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Core cloning Technique
    #7612055 - 11/08/07 05:04 PM (16 years, 2 months ago)

This is something ive been experimenting with. This is my second attempt.
I took a fresh Grifola frondosa i found in the wild...i put about 3 cc sterile water in an 18 guage syringe to provide backpressure, the dipped the needle in peroxide..and then stabbed it into the stembutt of the grifola. i removed it...cleaned the needle with peroxide and shot it into an LC made with karo.
here are the pics after 6 days







i made the LC of the "red blob" (photos in my journal and gallery) the same way...i didnt use peroxide that time because i feared the core sample being so narrow- perhaps the peroxide would just kill it. I have two LCs looking like the one in the pic- ill go to some grain soon and see if i can coax a contam...but so far it looks good!!


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Offlinefalcon
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Re: Core cloning Technique [Re: greys]
    #7612107 - 11/08/07 05:21 PM (16 years, 2 months ago)

Nice technique, it sounds like it might be helpful when trying to get a clean culture on agar when there is mold on the plate.


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OfflineRogerRabbitM
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Re: Core cloning Technique [Re: falcon]
    #7612207 - 11/08/07 05:56 PM (16 years, 2 months ago)

The biggest problem I have cloning wild fruits in my area is bacteria. Even with a core culture that has never been exposed to air, bacteria seems to be embedded throughout the fruit. How will you be able to isolate from bacteria in LC?
RR


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Invisiblegreys
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Re: Core cloning Technique [Re: RogerRabbit]
    #7612255 - 11/08/07 06:12 PM (16 years, 2 months ago)

Quote:

RogerRabbit said:
  How will you be able to isolate from bacteria in LC?
RR




I...have....no....idea:dancing:
  I dont have a flowhood yet and dont do much with agar...life would be so much easier for me if I did. Ive done something similar to this with wild white oysters and had about a 80% success rate.Oysters, are very forgiving, and this is new for me ,though. I found a couple grifola on halloween night in good shape so i decided to try some primitive tissue culture.
Im just excited it grew!:yesnod:

edit:pour speeling


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:greys:


Edited by greys (11/08/07 06:14 PM)


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OfflineFreeSporePrints
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Re: Core cloning Technique [Re: greys]
    #7613331 - 11/09/07 12:00 AM (16 years, 2 months ago)

Hi mates, hi greys!

Like Roger said the problem are the bacteria. This damned penciled organisms can bloom like rabbits (have you inoculed yet the grain jars?)..the only way to clean a contamined culture is to inocule a Petri plate with a drop of LC.

Why do not so starting directly from the Petri plates? I'm not sure but if i remember well Morelman said that he used to clean,with positive results, a contamined Morel LC using peroxide or chroline. Try to contact him.

And yes..i'm quite sure could be a problem of areas..in some areas the bacteria can be more then others..

Do you take the tissue from the inside of the fruitbody or inserting the needle from the external surface?

Thanks,
Fabio:mushroom2:


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