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Offlinemaster_yoshii
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Registered: 05/18/15
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What am I doing wrong?
    #21709697 - 05/22/15 09:09 AM (8 years, 8 months ago)

I just inoculated ten more jars last weekend and they are growing mold already! Little green mold is starting to show at the inoculation points within the verm/BRF mix. My very first two batches I ever did turned out perfectly, matter of fact, in about a week the cakes will be ready to put in a fruiting chamber. But the last two weekends all my inoculations have ended up moldy!!! Here is how I've been doing things:

I don't have a pressure cooker so I sterilize my jars for 90 minutes in a huge pot (steam sterilize)
I've been poking approx 4 holes in the tops of the jars as inoculation points.
I always sterilize my glovebox, matter of fact i wash it down in bleach every time before I use it.
I always use gloves and alcohol to wipe everything down before I touch it or use it.
I've been sterilizing the syringes before each inoculation into each jar. 
I store my jars uncovered on top of a bookshelf in my living room
The first 9 jars turned out beautifully, but now every jar since then has just gotten moldy, I repeat the process the same every time, and now i'm trying to figure out wtf is going on and why I'm getting mold. Any ideas? Anything I should try differently?


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OfflinePinPornProducer
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Re: What am I doing wrong? [Re: master_yoshii]
    #21709757 - 05/22/15 09:23 AM (8 years, 8 months ago)

Silly question but are you poking holes after you sterilize the jars?

Do you make your own syringes? If so what's the process?

And is the syringe you've had success with different than the one your using now?


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OfflinefirstTIMER420
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Re: What am I doing wrong? [Re: master_yoshii]
    #21709766 - 05/22/15 09:27 AM (8 years, 8 months ago)

I agree, when are you making the holes before or after sterilization?  and also, when you first got the contam, did you open the jars that were contamed before sterilizing them?  having all those spores released in your house might explain why everything you do no matter how sterile is getting re infected?

just my two cents


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Offlinemaster_yoshii
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Re: What am I doing wrong? [Re: PinPornProducer]
    #21709918 - 05/22/15 10:14 AM (8 years, 8 months ago)

I poke holes before i sterilize the jars and then cover them with the tape and also cover the tops with foil during sterilization, I remove the foil after the jars cool and then I put them in the glove box, the tape is only pierced once I inoculate them.

I did not make my own syringes. I purchased them from a store.


I have used different strains for each inoculation. The first two strains I used on my very first 9 jars which turned out wonderfully. All other strains (I've used 4 different ones since then) have all gotten moldy.


Edited by master_yoshii (05/22/15 10:22 AM)


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Offlinemaster_yoshii
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Re: What am I doing wrong? [Re: master_yoshii]
    #21709927 - 05/22/15 10:16 AM (8 years, 8 months ago)

I have no opened any jars that have had mold. I have thrown every one of them away. I don't even use them again or re-sterilize them, I just ditch the whole thing.


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OfflinePinPornProducer
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Re: What am I doing wrong? [Re: master_yoshii]
    #21709931 - 05/22/15 10:18 AM (8 years, 8 months ago)

What's your incubation temp?

Could be just really dirty syringes, have you looked into using agar yet?  Its fairly easy to get into and once you go agar you will never look back.


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OfflinefirstTIMER420
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Re: What am I doing wrong? [Re: master_yoshii]
    #21709938 - 05/22/15 10:19 AM (8 years, 8 months ago)

take the name out, spores are for microscopy purposes only.

not trying to be a dick, I did the same thing and then someone told me as well.

and from what your telling me I would think that you have mold spores in your house, I would give it a good scrubbing/cleaning before trying again.  also, water might be leaking in the jars during sterilizing and causing the contams, from my experience some water leaking is good, a lot is bad


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Offlinemaster_yoshii
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Re: What am I doing wrong? [Re: firstTIMER420]
    #21709964 - 05/22/15 10:26 AM (8 years, 8 months ago)

Yeah thanks, I removed the name. I'm new at this and microscopy interests me.

I do put tin foil over the tops of the jars when I sterilize them. Maybe it might be just a bad batch of spores?

I've been extremely careful and methodical in my sterilization and inoculation procedures.


I am open to trying out Agar. Where is a good place to buy it?


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InvisibleMad Season
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Re: What am I doing wrong? [Re: master_yoshii]
    #21710013 - 05/22/15 10:41 AM (8 years, 8 months ago)

Another silly question but not really. Do you have a dry verm layer? If you do, my guess is the syringes. Agar can be found at a grocery store in the Asian aisle or even better at an Asian market. It's usually a product called agar agar and you gotta ask nearly half the store to find it. Trust me though it's there.


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