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OfflineUrist
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Contaminant Isolation
    #21390746 - 03/10/15 11:31 PM (9 years, 2 months ago)

So I have bits dried mushroom gills and dust that may contain spores. I want to start these on agar but there will obviously be some contaminants. My question is how do I isolate the healthy tissue from the molds and bacteria and what equipment do I need to accomplish this effectively? Right now I just have the agar and some glass petri dishes.

I already know this will be difficult and is definitely not the best way to start out, but I don't feel like jumping through all the hoops and red tape necessary to get spore prints or syringes in CA.

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Invisiblewowimflabbergasted
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Re: Contaminant Isolation [Re: Urist] * 1
    #21390753 - 03/10/15 11:34 PM (9 years, 2 months ago)



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OfflineUrist
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Re: Contaminant Isolation [Re: wowimflabbergasted]
    #21390783 - 03/10/15 11:50 PM (9 years, 2 months ago)

Thank you.


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Offlinemushpunx
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Re: Contaminant Isolation [Re: Urist] * 1
    #21390879 - 03/11/15 12:31 AM (9 years, 2 months ago)

You might be able to swab a few spores out of them but I do know its entirely possible to revive dried mushroom tissue to get a culture going on agar.
I feel like if you were to just drop some gill dust on agar you would have too hard of a time cleaning away from contams

RogerRabbit said:

"Here's a process that has been 100% successful in reviving dry mushroom tissue for me so far. Don't use peroxide, whatever you do. It's toxic to fungi cells, and the last thing you want to do when reviving tissue is harm it further.

Sterilize several jars of distilled water, one of which should be prepared with small air exchange holes in the lid and a method of filtering, such as tyvek or synthetic filter disk, polyfill, etc.

Break up the dry tissue into grain sized chunks and put a dozen or so into one of your sterilized jars of water. Shake well to wash the pieces of tissue, and then transfer them to the next jar of sterile water. (you can pick up the pieces with a syringe needle or scalpel) Shake again to wash and repeat.

When you place them into the final jar, which is filtered, swirl them around in the water and then set the jar away and forget about it for a few weeks. You'll begin to see mycelium growing on any chunks that are viable. Don't use nutrients of any kind, just plain sterile distilled water. Once mycelium begins to form, transfer to agar to isolate away from any contaminants that may be present.
RR"

  Here I dug this up for you. Other people might chime in about this but this seems like the best way to go if you ask me, you would have a better shot startinh from live myc IMO

-mushpunx


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OfflineUrist
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Re: Contaminant Isolation [Re: mushpunx]
    #21390933 - 03/11/15 01:00 AM (9 years, 2 months ago)

Oh wow that seems pretty easy. For the sterile water I just need to boil the water and then put the jars in it for a bit to also sterilize them and then just seal them and stick a polyfill plug in the last one right?


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Offlinemushpunx
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Re: Contaminant Isolation [Re: Urist] * 1
    #21390938 - 03/11/15 01:03 AM (9 years, 2 months ago)

Quote:

Urist said:
Oh wow that seems pretty easy. For the sterile water I just need to boil the water and then put the jars in it for a bit to also sterilize them and then just seal them and stick a polyfill plug in the last one right?





When I sterilize water I run it through a pressure cooker cycle, not boil. You would have to havr the GE filter on the last jar before you pressure cook it, polyfill would probably work but maybe look at some of the GE lids designed for liquid culture they might be better suited for this

Good luck!


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