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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Inoculated my first jars. Don't feel good about them...
    #11006331 -

Today I made syringes and inoculated my first jars.  I have a feeling I'm going to have problems, but I suppose we'll see.  To begin,

Making the substrate went great.  I used the PF Tek, and was pleased with the substrate I produced (it would take impressive amounts of incompetence to mess this step up).  I scooped it into tall half-pint jars as I couldn't find the short-with-big-diameter sort.  I figured this was fairly trivial, but it did annoy me somewhat.

Sterilization of the jars went well.  I managed to successfully use the pressure cooker without burning myself.  I was feeling pretty good at this point.  I let it cool over dinner, then came back to make the syringes...

I alcoholed a shot glass and boiled it, then drew boiling water into the syringes.  Let everything cool in a really crappy glove box, then began scraping spores into the glass, sterilizing needle, spraying in, drawing up, etc.  I made 9 this way, as I destroyed one on accident with the flame sterilizing process...  I feel bad about this entire step. If I do get contams, I'm pretty sure this will have been their origin.  I had trouble manuevering the print, my gloves got sweaty and I couldn't hold things well, my glove box sucked, there were just too many variables here.  I was rather generous with spores, so I'm hoping I might save myself there.

Then there was inoculation.  I found out my needle points were too short to pass the layer of dry vermiculite. I felt really stupid, and did the best I could with the short needles, shaking the jars afterwards in hopes that at least a couple [thousand] spores would hit actual substrate.  Oh, and I had a ton of trouble removing the cooked on micropore tape, so I wound up shooting through it, then retaping the holes.  This didn't feel right either.

So I made 5 jars, and I'm hoping for at least 1 successful colinization, but expecting 0.  I'm soaking some birdseed, so I'm going to try my hand at that method tomorrow with my remaining syringes.  I think I'd have been better starting with a preloaded syringe, but I suppose this is how learning works. 

For kicks and giggles, I made up some more PF substrate, and scraped my sporeprint dry (I figured it was too contaminated by this point to be worth saving) directly into it, and mixed without sterilization or anything.  Then I buried the whole mess of it in my hobby greenhouse.  I'm sure the Universal Law of Irony will ensure that this is the only thing that fruits v_v  Maybe I should stick to plants. They're easy.

Any comments, suggestions, and protips are welcome.


--------------------
The Plant Mage Guild

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Re: Inoculated my first jars. Don't feel good about them... [Re: Horticulture]
    #11006345 -

I was rather generous with spores, so I'm hoping I might save myself there

Using more spores will work against you, Higher chance to contaminate actually


--------------------

Pf tek done right
How to Sterilize a Syringe
Quick and Easy Pasteurization
Quick and Easy LC
Pan Cyans without a PC
4 Easy step rye

AMU Q&A link

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Re: Inoculated my first jars. Don't feel good about them... [Re: Shea25]
    #11006476 -

It's definitely a learning experience.

Don't sweat it man. Vermiculite and BRF is cheap.

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Re: Inoculated my first jars. Don't feel good about them... [Re: Shea25]
    #11007945 -

Shea25 said:
I was rather generous with spores, so I'm hoping I might save myself there

Using more spores will work against you, Higher chance to contaminate actually


Please explain the logic in this?


--------------------
The Great Mycelium (TGMM) is more than you and me, we are all part of One.

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Re: Inoculated my first jars. Don't feel good about them... [Re: explosiveoxygen]
    #11008224 -

I like my syringes a little dark myself, so I am not sure what that poster means.  The only negative I know of about too many spores is clogging the syringe needle with too many spore clumps.

An easier way to make a syringe from a print is just to drop the print in a plastic baggie (inside a GB), inject water, mix it up a little, draw the spore water back into the syringe.  And you can just make more syringes with that one if you want.  That is how I do it anyway, and it has worked for years.

As far as the short needle, don't sweat it too much.  Sometimes the spores will germinate in the vermiculite layer and work their way down into the cake.  You can press the needle tip on the side of the jar and very slowly inject spore solution against the glass to drip down for a deeper inoculation point. 

But it is all about learning, and you know a heck of a lot more than you did just a few days ago.

Give those jars at least two weeks before you decide they didn't work.  Things can happen slowly some times.

Good luck.

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Re: Inoculated my first jars. Don't feel good about them... [Re: MLBjammer]
    #11008243 -

i scrape all the spores into a pile with my scalpel in a glove box then take a little scoop and drop in the back of the syringe and draw some pre sterilized water in it. i thought this would be disasterous at first, but i have a really low contam rate doing it like that.

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Re: Inoculated my first jars. Don't feel good about them... [Re: inkmaggot]
    #11008247 -

if there are too many spores then there is too much competition between spores to colonize, which can slow down colonization time thus leaving the door open a little longer for contams to enter.


--------------------
nappy then, nappy now, nappy for a bit
knee deep head over heals in this country shit

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Re: Inoculated my first jars. Don't feel good about them... [Re: explosiveoxygen]
    #11008848 -

Using more spores will work against you, Higher chance to contaminate actually


Please explain the logic in this?

In case the Print was contaminated or partially, by using lots of spores it greatly increases the chance of contamination. And also using loads of spore per syringe can slow things down and work against you also increasing the chance of contaminations taking hold


--------------------

Pf tek done right
How to Sterilize a Syringe
Quick and Easy Pasteurization
Quick and Easy LC
Pan Cyans without a PC
4 Easy step rye

AMU Q&A link

Extras: Filter Print Post Top
Re: Inoculated my first jars. Don't feel good about them... [Re: Shea25]
    #11008893 -

The glovebox is a lot more forgiving that you'd think. Based on what you wrote above I think there is a very high probability of success for the jars.

If you have contam, it would probably be from contams on the prints, not your procedure.

piercing thru the micropore tape is a non-issue because the dry verm is all the barrier you really need. I know u said u got some liquid in the dry verm, but when you gave the jars a little shake you probably scattered enuff dry verm across the top to create a barrier. The double layer of micropore over the holes might reduce your gas exchange later. But I'm bettting you're going to be just fine.


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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