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Offlinemothballs
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cloning, am i doing this right?
    #23348961 - 06/15/16 09:43 PM (7 years, 7 months ago)

so, i carved out the pith of the stem a fresh heathy mushroom in a glove box, blended it in a sterilized blender, sucked it up in syringes, and inoculated jars( which didnt grow) and a 4%honey water lc (still waiting).... am i doing this right?, is agar 100% necessary? any suggestions?


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Invisibletito123
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Re: cloning, am i doing this right? [Re: mothballs]
    #23348974 - 06/15/16 09:46 PM (7 years, 7 months ago)

Seems like sticking the mushroom tissue in a blender wouldn't be very sterile


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OfflineCaptcrunch90
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Re: cloning, am i doing this right? [Re: mothballs]
    #23348985 - 06/15/16 09:49 PM (7 years, 7 months ago)

I mean agar isn't 100% necessary, but you're also not 100% guaranteed to get a clean culture from the stipe. If you use that culture to make the LC and it was contaminated, well now your LC is contaminated. And if you don't test out your LC on a test jar, well then you've just ruined a bunch of grain jars.

Or you can just clone to agar and visually inspect/see any contamination and possibly save yourself a buttload of heartache and time. It's a gamble, but it's up to you man.


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Offlinemothballs
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Re: cloning, am i doing this right? [Re: Captcrunch90]
    #23348995 - 06/15/16 09:53 PM (7 years, 7 months ago)

thanks,... so in theory, blended pith in an lc would work assuming there are no contams


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OfflineCaptcrunch90
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Re: cloning, am i doing this right? [Re: mothballs]
    #23349016 - 06/15/16 09:59 PM (7 years, 7 months ago)

I only know about taking a biopsy with a syringe (bad idea), and agar as I explained. Couldn't tell ya man to be honest.


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Invisibletito123
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Re: cloning, am i doing this right? [Re: Captcrunch90]
    #23349569 - 06/16/16 02:06 AM (7 years, 7 months ago)

You blended the pith in a liquid culture solution?

It's be very risky to do in the first place since it's a harder, unnecessary step to ensure the blender is sterile and if everything went smoothly, you'd still want the mycelium to grow through the liquid culture before innoculating jars or another lc


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Offlinemothballs
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Re: cloning, am i doing this right? [Re: tito123]
    #23349591 - 06/16/16 02:16 AM (7 years, 7 months ago)

no, i premade the lc,sterilzed it and injected the blended tissue culture into the premade & sterelized lc


Edited by mothballs (06/16/16 02:19 AM)


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