Techniques for isolating monokaryotic mycelium

Discussion in 'ADVANCED MYCOLOGY' started by FewTrueSeed, Nov 25, 2016.

  1. FewTrueSeed

    FewTrueSeed Many meiosis Supporter

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    In my journey to create crosses i have considered making mass multispore innoculation and cloning the outliers.

    However there is something exciting about having a monokaryotic culture to return to.

    The method i am familiar with involves diluting viable spores and sectoring out very early in germination. I have a micriscope to cheak for clamp connections.

    Does anyone have any experience or advice with this procedure?
     
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  2. PE-n-hed

    PE-n-hed Well-Known Member Moderator Mushroom Doctor Cannabis Doctor Supporter

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    In regards to crosses, I have no first hand experience .some more experienced members may chime in.
    I remember reading on it ,when Workman created APE, it's been awhile.
    - serial dilution
    - streaking agar and finding the last smallest colonies to avoid spore clumping
    -Finding no clamps as starting point
    You are headed in the right direction for sure.
    I know that The Chosen One has experience
    He is a active member ,but hasn't posted since 2015
    I can't help with the process ,but will support you:scope:
    Now as far as isolation ,I have done one from spore ,transfers from sectors till no more sectoring.
    I feel I just lucky that time.it was Texas and did great. All my other iso monocultures came from a ms grow ,cloned fruit bodies ,then further agar work till a strain was isolated.
    But it is my understanding that a culture from clone is not a true isolate .but for cultivation purposes is all I needed. I am by far a geneticist or lab guru
    I am always here to learn
     
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  3. GoldiBoi

    GoldiBoi New Member

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    So are you saying a culture from clone, as in, a culture from a fruitbody, is not a true form of strain isolation...how so? I did not know this
     
    Last edited: Jan 2, 2017