Techniques for isolating monokaryotic mycelium

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

  1. FewTrueSeed

    FewTrueSeed Many meiosis Supporter

    Sep 29, 2014
    Green Mountains
    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?
    GlassKrakr likes this.
  2. PE-n-hed

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

    May 31, 2011
    Planet X
    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 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
    treewood, redcat, muskie and 3 others like this.
  3. GoldiBoi

    GoldiBoi New Member

    Dec 23, 2016
    So are you saying a culture from clone, as in, a culture from a fruitbody, is not a true form of strain so? I did not know this
    Last edited: Jan 2, 2017
  4. SannyTIES

    SannyTIES New Member

    Nov 11, 2019
    A clone tissue sample contains what we call "a mosaic isolate" it contains 100s of strains in it. Ever notice how many sectors come from a single tissue sample? It's not a pure isolate. Not even close.

    A clone tissue sample will die out or senescence rather quickly. In other words cloning is a one way ticket to dead end. It can produce a next grow, but not the answer to creating a pure culture.