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OfflineIvan
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Better Mycelium syringes
    #2016910 - 10/17/03 03:16 AM (13 years, 8 months ago)

Mad scientists please give your advice :-)

AFOF has used the honey/water tek for liquid inocilum whith varying success. After these experments it was the advise of another FOF to use a PDA solution with 3 times the water content and inoculate with multispore/water solution ala spore syringe transfer.
I have had a few ideas of using some ground end substrate and adding this to the PDA mixture to help the mycelium adapt(P. Stamets GGMM).
Any great ideas out there?


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OfflineAnnoA
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Re: Better Mycelium syringes [Re: Ivan]
    #2016913 - 10/17/03 03:19 AM (13 years, 8 months ago)

Here, a different method:
http://www.shroomery.org/index/par/7943


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OfflineIvan
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Re: Better Mycelium syringes [Re: Anno]
    #2017483 - 10/17/03 10:43 AM (13 years, 8 months ago)

Thanks Anno,
I know and like the method you mentioned and have used it in the past.
If the equipment is available and you want to inoculate right away then its absolutely the way to go.
The question is developing the mycelium with a minimum of transfers.
Give you an idea, 1 x 20ml ampule of sterile water with injectable stopper is sucked into a syringe of the same capacity.
The spore print taken on a glass petri is then saturated with the sterile water and sucked into a new clean syringe.
Then, transferred back to the 20 ml ampule (neat trick ;-) )This gives a very concentrated solution of spores in a minimum size container.
This can be re-extracted to syringes containing medium. (ie mycelium growing in the syringe)
3 benifits of doing this.

1.) Mycelium is produced in its early stages thus growth is more vigerous. (this is my observation not a proven fact)

2.) Contamination vectors are reduced a the syringe itself is propagated.

3.) Being multi-spore inoculation, master syringes can be selected based on growth traits, and the addition of substrate to the medium can act as an acid test for substrate viability.

I remember Stammets stating that mushrooms have a type of genetic memory and that it will take a few generations of growth to adapt to a new substrate.
With this said I believe it's possible that a print taken from a "bird seed" substrate might be less productive on its first run
on rye grain (for example).



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Invisiblemicro
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Re: Better Mycelium syringes [Re: Ivan]
    #2017518 - 10/17/03 11:09 AM (13 years, 8 months ago)

You can use a septum plug on a mason jar, too, so there's a self-healing injection site, and culture 500 mls at a time.

If you want to culture inside the syringe make sure to leave room for some sterile air. Otherwise, this should work fine.

What Stamets is talking about only applies if you're growing multiple generations on one type of media. Mushrooms will grow on any good substrate from spores, or from liquid media made from the spores, but if you keep using liquid media to inoculate more liquid media after a while the strain will "degrate." This is because it loses a lot of the enzyme functioning that may be important for another substrate but not for the liquid media. One or two generations in liquid culture is fine, though.

--
Micro


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OfflineIvan
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Re: micro [Re: micro]
    #2017615 - 10/17/03 12:17 PM (13 years, 8 months ago)

Micro,
Execelent idea about the septum plug on a mason jar and I will agree with you on generational mycelia degradation in liquid inoculate.
but what I am proposing is to add the substrate to the liquid/semi-liquid media for the reason of adaptation and proper medium selection.
Some of the top selling mycological agar include fine ground grains and nutrients for this reason.
What I have found from my own experiences is the following.
In the past I have used rye grain, bird seed, pure millet, enriched millet, ect...

What you say is true, that all of these substrate mediums are fine for growing a range of mushrooms, the spore prints I made always grew better on the same type of substrate the original print came from.
It could be a fluke but I tend to believe that this is not the case.
I say this because as the spores germinate and create hyphae this primary trigger is evironmental (nutrients, medium, climate,).
By giving them an extra boost in the nutrients, one is thus adapting the genetic tolerance for a given substrate.

I, in no way, claim to be right.. Just an observation





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Invisiblemicro
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Re: micro [Re: Ivan]
    #2017697 - 10/17/03 01:08 PM (13 years, 8 months ago)

Two each their own. I get great growth on PDA, but I also supplement it with amino acids. Agar won't have actual grain in it, but more the hot-water soluble extract from the grain. Nutrients help in agar up to a point, but you don't want it to be too saturated with stuff or you'll get repressed and inhibited growth. Also keep in mind the C:N ratio should be about 10:1.

No reason it couldn't work -- you may just want to do a hot-water soak on some grain and use the water to make the agar (along with potatoes if using PDA,) -- you want the agar to be homogenous. Maybe just reduce the amount of potatos a bit.

--
Micro


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(Avatar is Maxxy, a character by Mizzyam, RIP)


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Offlinelysergic
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Re: micro [Re: micro]
    #2044357 - 10/26/03 11:19 AM (13 years, 7 months ago)

I'd recommend a simmer of WBS, cracked corn, some compost, and taters. Simmer for an hour or so after a 24 hour soak, then use that liquid. If you wanted a super-concentrate, get that liquid, then boil some of the water off.


The reason I recomended grains, poo, and taters, is that you are going to keep whatever enzymes are needed for growing on all of those cultures. Maybe mix in some karo with this, then sterilize. Not sure if it would work or not, just my thoughts. It would also be very very cloudy, and prevent ou from seeing contams. However, after you get to the point that you are making 5-10 karo syringes a week with no contams, it shouldn't be a big issue.


--------------------
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PsiloKitten said:
Just give em a little more time, the iraqis are making great progress. And this is unorganized. Wait till they get organized.


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Invisiblemicro
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Re: micro [Re: lysergic]
    #2045202 - 10/26/03 06:09 PM (13 years, 7 months ago)

If you only need inoculum and have a bag of colonized substrate, here's a good trick:

Inject lots of sterile water into the bag down into the corner of the bag (keep in ming the mycelium is going to suck up the water, so don't let it touch it too much)

Cruble a chunk of mycelium and mix it with the water in the bag.

Sterilly draw this mycelium water into syringes.

This way is virtually contaminant-free....

--
Micro


--------------------
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(Avatar is Maxxy, a character by Mizzyam, RIP)


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