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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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My clones contam
    #25956926 -

Ok so I made agar, done transfers and all that good stuff and nothing every contams. So I feel confident in my SAB Tek. But every time I try to take a clone of the biggest booms I grew it always turns slimy on me and smells fermented. How can I beat the contam and keep my genetics.? Plz help. Thx

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Re: My clones contam [Re: Hobbit GDF]
    #25956947 -

Use small chunks from the inside of the stipend. Hold yours plates/jars upside down/ vertically with an upside down tilt with your lid in vertical orientation when performing tranfers. Take the leading edge and only a piece the diameter of a ball point pen body. If contamination persists transfer to a plate containing a 10 percent solution of H202 before transferring to a clean dish. Don't make an LC or grain master from your peroxide dish, move it one last time to be positive.


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My no pour Petri TEK - https://www.shroomery.org/forums/showflat.php/Number/27252059/page/1

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Re: My clones contam [Re: Professor X]
    #25956959 -

Walk us through your biopsy method. My guess is that you're cutting into the stipe instead of pulling it apart or not flame sterilizing.


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"I members.... do you members"
Memberberries circa 2016


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Re: My clones contam [Re: Psilo_citizen]
    #25957018 -

Ok so I set up SAB like I'm supposed to. I have it loaded with what I need. Torch and scalpel next to it handy. I soak hands in alcohol and go pluck the lucky Boomer. I put it in a sandwich baggie for transportation. Clean the outside of bag and load it in SAB. I let it settle for few minute. I torch the knife and enter the sab. I hold the knife. I never sit it down. I get the specimen out of bag. Rip it open along the stem with my fingers. I use knife to scrape a little piece out and open my agar dish and try to get it off into center of agar. The close it up.

I've done spore inoc on agar and a2a transfers and a2g without an issue of contam. So it must be the specimen right? I hope I was accurate in my walk through.

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Re: My clones contam [Re: Hobbit GDF]
    #25957028 -

Hobbit GDF said:
Ok so I set up SAB like I'm supposed to. I have it loaded with what I need. Torch and scalpel next to it handy. I soak hands in alcohol and go pluck the lucky Boomer. I put it in a sandwich baggie for transportation. Clean the outside of bag and load it in SAB. I let it settle for few minute. I torch the knife and enter the sab. I hold the knife. I never sit it down. I get the specimen out of bag. Rip it open along the stem with my fingers. I use knife to scrape a little piece out and open my agar dish and try to get it off into center of agar. The close it up.

I've done spore inoc on agar and a2a transfers and a2g without an issue of contam. So it must be the specimen right? I hope I was accurate in my walk through.


Sounds pretty good, but the bag is unnecesary. You're doing basically exactly what RR recommends you do:



I think we'll probably need some pictures of the bad results to help troubleshoot it further.


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My Shoebox Factory Beginner's Tek | Shoebox Assembly Tek | Methodology/teks I use | How I Store Bulk Items

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Re: My clones contam [Re: ShaperDreaming]
    #25957042 -

Yeah I'll do that later. I need to do some transfers tomorrow so I'll take pictures. Stay tuned.

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Re: My clones contam [Re: Hobbit GDF]
    #25957105 -

Last week I did clones with my wife, made it ridiculously easy. She tore it open and I sliced the samples out. I take 3 just to be sure. Some of our processes are 100x easier with a helper. If you don't have a trustworthy helper get an adjustable bench clamp, they sell them at Hobby Lobby and Wal Mart for working on models.


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My no pour Petri TEK - https://www.shroomery.org/forums/showflat.php/Number/27252059/page/1

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Re: My clones contam [Re: Professor X]
    #25957174 -

Meh...
I'd much rather do sterile work solo. Every additional hand is an increased risk for contamination regardless of added convenience. Besides, with decent technique, grabbing a tissue culture really shouldn't be a two person job.


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"I members.... do you members"
Memberberries circa 2016


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Re: My clones contam [Re: Psilo_citizen]
    #25957193 -

My wife helps me when I need her. It makes it easier for sure. Love my wife.

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Re: My clones contam [Re: Hobbit GDF]
    #25957210 -

You guys working in front of laminar flow I take it?


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"I members.... do you members"
Memberberries circa 2016


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Re: My clones contam [Re: Psilo_citizen]
    #25957456 -

Psilo_citizen said:
You guys working in front of laminar flow I take it?


OP mentioned an SAB.

I suggest trying any or all of the following:

  • Wear gloves and a dust mask when you select the specimen and the entire time you clone
  • Flame your blade before taking the specimen and after every contact with the tissue
  • Look up 9er tek's method of peeling stem tissue like a banana
  • Dedicate a few jars to invitro fruiting for the purpose of cloning, try cloning tissue from one and pins from another
  • Dedicate one or more shoeboxes/containers/whatevers to the same, keep them as clean as possible during spawning and consolidation and then try cloning some pins when they come in

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Re: My clones contam [Re: SpikeSpiegel]
    #25957538 -

Quote:
OP mentioned an SAB.



That was directed at professor X, not op.

There's zero need to have a special clean grow for cloning. There's likely an error in his technique and be needs to figure out what that error is, not create crutches to avoid the problem. Spawning is an inherently unclean process anyway.


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"I members.... do you members"
Memberberries circa 2016


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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