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OfflineDr.Sparky
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Registered: 10/06/23
Posts: 19
Last seen: 4 hours, 22 minutes
Needle Gauge, Tissue Sample
    #28494186 - 10/06/23 07:47 AM (3 months, 21 days ago)

Working with my grandson on growing some oyster mushrooms.  Our plan is to take a needle biopsy of a store bought mushroom and inject it through an Injection port into sterilized liquid medium.  After mycelium growth we would inject 10 ml into a sterilized bottle of prepared rye berries and finally mix the berries into a 5 gal bucket of straw or wood pellets.
My question is regarding the needle: is 18 ga too small vs 16 ga?  Also, will a blunt needle destroy the rubber port?

I am concerned about contamination and feel that pressure cooking the liquid medium, the rye berries, etc,  will give us the least exposure to contamination.


Edited by Dr.Sparky (10/06/23 07:50 AM)


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OfflineDharmaForKarma
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Registered: 04/24/20
Posts: 565
Last seen: 1 day, 7 hours
Re: Needle Gauge, Tissue Sample [Re: Dr.Sparky]
    #28494199 - 10/06/23 08:07 AM (3 months, 21 days ago)

Welcome!

Yes. There is a great tek on this very thing. It works quite well with cubensis. Someone here or in the Gourmet section may known if it’s appropriate for oysters.

Josex' Poke: A different way of doing LC
https://www.shroomery.org/forums/showflat.php/Number/24740168


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InvisibleQenwi
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Registered: 04/15/23
Posts: 10
Re: Needle Gauge, Tissue Sample [Re: Dr.Sparky]
    #28494421 - 10/06/23 01:02 PM (3 months, 21 days ago)

You can use either 18 or 16 g. I'd recommend splitting the mushroom and not touching the middle of the stem just to make sure you're taking a sterile tissue. The problem with store bought mushroom is you don't know if the mushroom mycelium generation is new or old. So you shouldn't depend on it for bulk production. Also, try to take the tissue with sterile needle and transfer it to agar plate just to make sure it's 100% clean. Then take a small wedge of mycelium to make another transfer to sterile liquid medium


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OfflineDr.Sparky
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Registered: 10/06/23
Posts: 19
Last seen: 4 hours, 22 minutes
Re: Needle Gauge, Tissue Sample [Re: Qenwi]
    #28494587 - 10/06/23 03:58 PM (3 months, 21 days ago)

Luckily we have a local grocery that gets delivery of mushrooms from a farm that cuts them the day of delivery.  Each week they may differ but they are super fresh.


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InvisibleCrayolaHalls
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Registered: 03/23/10
Posts: 216
Loc: The Lot
Re: Needle Gauge, Tissue Sample [Re: Dr.Sparky] * 1
    #28497094 - 10/08/23 08:49 PM (3 months, 18 days ago)

Quote:

Dr.Sparky said:
Luckily we have a local grocery that gets delivery of mushrooms from a farm that cuts them the day of delivery.  Each week they may differ but they are super fresh.




I think what Q was referring to was the generation of mycelium at the producer.  Multiple transfers of mycelium.  There is a limit to the number of times you can clone a clone.  You should be fine, but if you plan to grow for a while using that store sample you should plan to eventually take spores from your production and start a new line.

But definitely, yes, the fresher the better when taking a sample from the store.


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