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Invisibleohmatic
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Agar Wedge / Mushroom Tissue to mycelium syringe - ohm's ghetto tek
    #3495897 - 12/14/04 06:21 AM (12 years, 5 months ago)

hey!

so ive had this nice plate of colonised agar i decided to play around a bit
and make a mycelium syringe, why not.

what ull need:
1.) a lil jar with a screw on lid, with a polyfill/tyvek(better) filter
2.) syringes with some big gauge needles
3.) scalpell
4.) h2o2
5.) lighter
6.) glass sheds
7.) plate of colonised agar :wink:

1.) soo u get ur goods together, fill the lil jar with about 1/5th of it volume with
water, throw in the glass sheds, clean the syringed and the scalpell and
pepare it all:


2.) next thing to wrap it all nicely in foil and load to pc and pc for 30 mins or more:


3.) let cool till cool :wink: - in my case about 8 hrs.

4.) clean ur workspace in the meantime, spay it down with lysol and
unload the goods.

5.) quickly cut a wedge of the plate, minimally open the jars lid (which u didnt
screw on tightly in the beginning so u could easily remove it in this step),
inject about 5cc of h2o2 (30% used here)
and start to shake.

here a pic of the workplace with the wedges already int he jar:


6.) now, if u used tyvek under the lid, ull be able to shake it like a mad man
getting MUCH better results than i did, since i could just let it circle
since otherwise the suspension would come through the polyfill...

anyway u end up with this:


7.) draw to syringe and  nock up jars.

--

well whats happening is basically, the glass sheds will rip off some tissue of the
wedges, the h2o2 will grill eventual contams and there u are,
nice mycelium syringe the easy way.

uhm, yeah, i knocked up 6 jars to see if this was any good,
the liquid in the syinge looked rather milky and i saw some stuff floating
around in it, hopefully myc and not just shredded agar.

oh yeah, and i GREATLY advise the use of tyved over polyfill,
sicne it really sucked not being able to shake it like a madman but having
to care about nothing oozing out the polyfill, with tyvek ull be able to go
just so much berserk that u dont kill the tyvek :wink:

hope that was usefull.
peace ohm :mushroom2:

*edit*
if anyone wonders why i wrote this all up,
it was the intention to make everyone able to do mycelium syringes
without fucking expensive devices like eberbach containers or somethign.
in the theory it worked real nice,
lets see how colonistion will be like.

*edit2*
fixed the wrong ammount of h2o2, i checked the syringe again and found
it was only 5cc.

importand edit

i just realised something.
actually, that procedure should be 'useable'
for direct cloning, say, instead of the wedge throw a piece of mushrooms
tissue into the lil jar, shake as writen and suck up the shredded mushroom
directly !!!

i will try and report back to u!


Direct Mushroom Tissue to Mycelium Synringe

allright, so, since ive been thinking a bit more about the tek i posted
i want to present its second great ability, to directly clone mushroom tissue.

for this reason there will be some modifications.

Things that will be done:

now the lil jar is filled with 12cc of water and more glassheds.
beneath the lid, a square of polyproplylene plastic is put and the lid
is screwet on lightly.
there is no filter in the hole.
its screwed on lightly (remember not tight to prevent builup of
vakuum upon cooling)
and alu wrap is fold over it, secured with a rubberband.

now all is getting pressurecooked.

what will happen next:

after all cooled, the aluwrap will be reoved and the cap with the
polypropylene square minimally liftet and a piece of extracted mushroom
tissue will be inserted.

now the cap is screwed on tightly and the crap is being shaken out of
it.

the glass sheds will rip the mush tissua apart and spread it in the sterile water.

next, a sterile syringe is drawn up with 1cc of 30% h2o2 and injected
through the hole in the lid, LEAVING the needle in the hole (since it will
pierce through the polypropylene square)

and the water/myc mix is gently shaken around to evenly distribute the injected
h2o2 to grill whatever there may be.

next, syringe is drawn up with the shredded tissue and h2o2 water.

finally, nock them jars.

heh sounds sweet dont it? well, lets see if it works out the
way it does in my fantasy :wink:
peace ohm :mushroom2:

Allright i did the Direct Tissue Cloning
as described above, i proceeded and shredded a piece of straight inner
tissue from a mushroom.

heres the final product:


oh and jeah, thats the guy that was cloned:


as u can see, there is noticeable sediment / mushroom mycelium flakes
in the syringe, i used it to nock up 3 wbs jars,
if it all works out i should see growth within some day.

comeone i wanna hype up my tek :wink:
peace ohm :mushroom2:

*edit*
allright it took a long ass time but a week ago i noticed mycelium
growth on 4 jars.
soo the whole thing actually worked out :smile:
i cannot wait to see how things will proceed, its the first liquid inoculant / clone
ive ever done :laugh:

sooooo cool!
peace ohm :mushroom2:


Edited by ohmatic (01/11/05 06:50 AM)


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InvisibleMagashM
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ohmatic]
    #3495901 - 12/14/04 06:27 AM (12 years, 5 months ago)

:thumbup: :grin:


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OfflineAnnoA
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: Magash] * 1
    #3496037 - 12/14/04 07:34 AM (12 years, 5 months ago)

10ml of 30% peroxide for such a small amount? i hope this won't too much...

Here is how I do it for small amounts:

Take a colonized, clean petri dish and a syringe full of sterile water.


Inject some water on the agar surface and scrape the mycelium layer, then suck the mycelium fragments into the syringe.



Result:



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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: Anno]
    #3496120 - 12/14/04 08:13 AM (12 years, 5 months ago)

i just checked, it was just 5cc, ill edit the guide.
when should i see first signs of colonisation?
peace ohm :mushroom2:


--------------------
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InvisibleADoobie
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ohmatic]
    #3496264 - 12/14/04 09:53 AM (12 years, 5 months ago)

Good guid Ohm...but Anno has ya beet there. I was about to try your tek till anno spit that one out...

However, it looks like with Ohm's tek you can store your petri dish away and use it later...Anno, your method looks like you better toss what you aren't going to suck up in a syringe...Am I correct?


--------------------
ADoobie said:
I know what strikes me as odd about this. It is totally unnatural. The whole idea is just totally backwards.

So are historians going to look back at 'The Great Cake Eaters'? Damn. That was a backwards step in the evolutionary process. We left Shroomheads in the dust. Welcome to the new age of Vermheads.

VvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVv

I ONLY STUDY legal, edible mushrooms and mycology. Any mention of any other kind of mushroom is purely fictional, and pictures are ripped from the internet. I do not grow mushrooms!


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Invisibleohmatic
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ADoobie]
    #3496284 - 12/14/04 10:13 AM (12 years, 5 months ago)

Quote:

ADoobie said:
Good guid Ohm...but Anno has ya beet there. I was about to try your tek till anno spit that one out...

However, it looks like with Ohm's tek you can store your petri dish away and use it later...Anno, your method looks like you better toss what you aren't going to suck up in a syringe...Am I correct?




actually i expected someone to post about scratching mycelium of the
surface and sucking it up.

anyway, u dont have to discard the petri if u do it annos way,
only, if the water was unsterile ull have problems since u expose
the surface of the agar to the water.

so, if that was untersile and loaded with whatever, itll spread on the dish.
if not, u should be fine.

either or cant happen if done my way.
peace ohm :mushroom2:


--------------------
:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !


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InvisibleADoobie
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ohmatic]
    #3496307 - 12/14/04 10:29 AM (12 years, 5 months ago)

Good point...

Now I wonder if the larger bits of un-broken myc in anno's tek

a) will clog your needle
b) will be "more established" and speed up colonization?

Also, I wonder if the smaller bits of broken up myc in ohms tek

a) will allow for better solution dispersion, speeding up b)
b) will be "more evenly dispersed" and speed up colonization?

I guess this is not so much an "anno's idea vs ohm's idea", but is instead just taking a closer look at both...


--------------------
ADoobie said:
I know what strikes me as odd about this. It is totally unnatural. The whole idea is just totally backwards.

So are historians going to look back at 'The Great Cake Eaters'? Damn. That was a backwards step in the evolutionary process. We left Shroomheads in the dust. Welcome to the new age of Vermheads.

VvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVv

I ONLY STUDY legal, edible mushrooms and mycology. Any mention of any other kind of mushroom is purely fictional, and pictures are ripped from the internet. I do not grow mushrooms!


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OfflineAnnoA
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ADoobie]
    #3496308 - 12/14/04 10:30 AM (12 years, 5 months ago)

>I wonder if the larger bits of un-broken myc in anno's tek will clog your needle

They came into the syringe, they will come out....


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InvisibleADoobie
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: Anno]
    #3496322 - 12/14/04 10:39 AM (12 years, 5 months ago)

True...then again, I was thinking they went in a little at a time. I then assumed that when the needle points down for inoculation, the sediment will drift to the bottom, and all of it will try and flow out the needle at once...

But I think I get the drift...it's not gunna clog your needle.


--------------------
ADoobie said:
I know what strikes me as odd about this. It is totally unnatural. The whole idea is just totally backwards.

So are historians going to look back at 'The Great Cake Eaters'? Damn. That was a backwards step in the evolutionary process. We left Shroomheads in the dust. Welcome to the new age of Vermheads.

VvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVv

I ONLY STUDY legal, edible mushrooms and mycology. Any mention of any other kind of mushroom is purely fictional, and pictures are ripped from the internet. I do not grow mushrooms!


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Invisibleohmatic
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ADoobie]
    #3496327 - 12/14/04 10:42 AM (12 years, 5 months ago)

Quote:

ADoobie said:
True...then again, I was thinking they went in a little at a time. I then assumed that when the needle points down for inoculation, the sediment will drift to the bottom, and all of it will try and flow out the needle at once...

But I think I get the drift...it's not gunna clog your needle.




the clogging u referr to, IF at all, could just happen as much in mine,
as in annos way.

neither nore will really prevent it, but actually, by pressing the plunger
u should be able to shoot it out anyway.
peace ohm :mushroom2:


--------------------
:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !


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InvisibleADoobie
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ohmatic]
    #3496349 - 12/14/04 10:54 AM (12 years, 5 months ago)

So then the only question of mine that remains is,

Will chunks or watery solution colonize faster, or will the difference be negligible?


--------------------
ADoobie said:
I know what strikes me as odd about this. It is totally unnatural. The whole idea is just totally backwards.

So are historians going to look back at 'The Great Cake Eaters'? Damn. That was a backwards step in the evolutionary process. We left Shroomheads in the dust. Welcome to the new age of Vermheads.

VvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVv

I ONLY STUDY legal, edible mushrooms and mycology. Any mention of any other kind of mushroom is purely fictional, and pictures are ripped from the internet. I do not grow mushrooms!


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Invisibleohmatic
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Re: Agar wedge to Myc Syringe - ohmatics ghetto way [Re: ADoobie]
    #3496356 - 12/14/04 10:57 AM (12 years, 5 months ago)

my logic tells me, that the watery solution would colonise faster
since it can be on more grains at once, but this is just theory.

anyway, i just realised i could use that procedure for direct cloning
and since i just have a mush id like to clone,
ill try and also report back!

check the important edit in the initial post for the idea.
peace ohm :mushroom2:


--------------------
:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !


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InvisibleADoobie
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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: ohmatic]
    #3496363 - 12/14/04 10:59 AM (12 years, 5 months ago)

Well shit son get on it! :wink:


--------------------
ADoobie said:
I know what strikes me as odd about this. It is totally unnatural. The whole idea is just totally backwards.

So are historians going to look back at 'The Great Cake Eaters'? Damn. That was a backwards step in the evolutionary process. We left Shroomheads in the dust. Welcome to the new age of Vermheads.

VvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVvVv

I ONLY STUDY legal, edible mushrooms and mycology. Any mention of any other kind of mushroom is purely fictional, and pictures are ripped from the internet. I do not grow mushrooms!


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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: ADoobie]
    #3496400 - 12/14/04 11:19 AM (12 years, 5 months ago)

This tek is too anal for me, I rather just innoc some spores into karo
:laugh:


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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: kronnyQ]
    #3496629 - 12/14/04 12:43 PM (12 years, 5 months ago)

i got a question concerning tissue extraction from a mushroom.

i know i shall rip it apart and take a piece from the inside.
shall i hold this piece into 30% h2o2 prior to throwing it into the 'shredding jar'

i guess it cant be bad, just do it hm?
peace ohm :mushroom2:


--------------------
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RIP #cultivation! ....can't associate? well FUCK U !


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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: ohmatic]
    #3496656 - 12/14/04 12:53 PM (12 years, 5 months ago)

If done aseptically , no need, just transfer piece to agar filled petri and let it go .


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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: tripndicular]
    #3496693 - 12/14/04 01:04 PM (12 years, 5 months ago)

Quote:

tripndicular said:
If done aseptically , no need, just transfer piece to agar filled petri and let it go .




im talking about directly shredding the tissue of the shroom inside
the shredding jar... not talking about agar.

i guess ill just do it,
cant be wrong about it.
peace ohm :mushroom2:


--------------------
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RIP #cultivation! ....can't associate? well FUCK U !


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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: tripndicular]
    #3496756 - 12/14/04 01:18 PM (12 years, 5 months ago)

Don't put it into straight 30% h202..that could harm it.just throw it in the mix the same as you did the agar wedge...you should edit the step when you mention opening the jar lid you left loose and injecting the h202 and add to put the wedge into the jar..you never actually tell the person when to put it in. Most people wouldn't even notice and just know you throw it in but doesn't hurt to clarify. And are you talking 5ml of 30% h202 going in to the mix? Or are you talking 30% of the solution is 3% h202? If you don't mean your using a strength of peroxide stronger then the normal 3% then you should clarify that as well. If you are using 30% h202 then an exact amount of water should be used so people can convert that to 3% since most people don't order extra strength h202.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: scatmanrav]
    #3496934 - 12/14/04 01:52 PM (12 years, 5 months ago)

Quote:

scatmanrav said:
Don't put it into straight 30% h202..that could harm it.just throw it in the mix the same as you did the agar wedge...you should edit the step when you mention opening the jar lid you left loose and injecting the h202 and add to put the wedge into the jar..you never actually tell the person when to put it in. Most people wouldn't even notice and just know you throw it in but doesn't hurt to clarify. And are you talking 5ml of 30% h202 going in to the mix? Or are you talking 30% of the solution is 3% h202? If you don't mean your using a strength of peroxide stronger then the normal 3% then you should clarify that as well. If you are using 30% h202 then an exact amount of water should be used so people can convert that to 3% since most people don't order extra strength h202.




good points u got in there,
ill go edit after i had dinner.
peace ohm :mushroom2:


--------------------
:penis: MONOTUB tek :sun: HEATBOMB tek :penis:

RIP #cultivation! ....can't associate? well FUCK U !


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Offlinescatmanrav
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Re: Agar wedge to Myc Syringe - ohm's ghetto tek [Re: ohmatic]
    #3496985 - 12/14/04 02:08 PM (12 years, 5 months ago)

Dinner? Its 2 in the afternoon, its not dinner time! Its breakfast time....


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


Post Extras: Print Post  Remind Me! Notify Moderator
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Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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