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Cloning Pins Directly to Agar
    #5703541 -

A few years ago, I ran several experiments to prove/disprove stamets' theory that if you clone very young healthy pins, the hormones that stimulate pinning are active, therefore the mycelium that grows from those pins will in turn produce better flushes with abundunt primordia and fewer aborts. The tests were inconclusive, but in producing my mushroom instructional DVD, I've started up many of the old experiments in order to demonstrate them on the DVD. Here's a sneak peak. These pins were snipped off a healthy casing layer that was supporting a massive pinset. In a glovebox, flame sterilized tweezers were used to hold the pins, while a flame sterilized scalpel was used to cut them off just above the casing layer. Anybody want to give odds on if cloning this way will work? I keep hearing that pins will rot. Whatcha folks think?
RR


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Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703567 -

It probably will work...

I did something simular...but I cut the caps off.

and they cloned just fine.


Plus we all know you seem to have a knack to do things that seem impossible!~

:smile:



tc


--------------------
Laterz, Road

Who the hell you callin crazy?
You wouldn't know what crazy was if Charles Manson was eating froot loops on your front porch!

Brainiac said:
PM the names with on there names, that means they have mushrooms for sale.


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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703580 -

i bet it works.

i've missed aborts before, dunked, saw them turn black and nasty, then mycelium started growing on them.  (perhaps feeding in a "cannibalistic" manner, eating the dead mushroom tissue?)  so even if they "rot" i say it still goes.  i don't buy that the hormones present in the pins help anything.  it would be such a minute amount, and when you take out a mycelium wedge you're starting at the beginning again, i don't see how pinning hormones present in your original sample has anything to do with a mycelium sample that's going to go through a colonization run before fruiting again... i have heard of collecting a bunch of pins, pulverizing and making a solution, and using that solution to mist a casing, and while that's more likely to work IMO i still doubt it... you are cloning a strain that you know is capable of fruiting, so you'll see gains there, and i could see how it could be confused that the hormones did it, but i think any "prolific fruiting" experienced from the clone would be no more than a good choice in strain isolation

and what exactly is "rotting" anyway?  when we think of rotten food its usually obvious, covered in mold sometimes, and usually stinky (bacteria)... i don't see how a clean pin kept in a sterile plate could rot... but then again the likelyhood of a pin itself being sterile depends on your growing environment and isn't too likely... maybe washing them off in a strong h2o2 solution (and maybe even using antibiotic agar) would prevent that?  dunno i'm babbling... :smile:

that's an awesome petri btw, where does one find those with the nifty separators?

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Re: Cloning Pins Directly to Agar [Re: creamcorn]
    #5703617 -

what agar is that?

i wonder, if you use some sort of peroxide agar/antibiotic agar i see good chances that itll work.
given those pins do have bacteria on them or alike (after all thats why we take tissue from inside otherwise) youd need
some component to keep eg molds form growing before the pin myc takes over the agar.

curious hows it gonna work :smile:
god agar, if i knew how much fun it'd be id started 2 yrs ago with agar.


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

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

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703621 -

Cool looking dish.

I think it will work.

Why does a pin have better hormones than a cut out wedge?


:sun:


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Child of the 60's, Tripping ever since.


:sun:

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Re: Cloning Pins Directly to Agar [Re: ohmatic]
    #5703642 -

I dont know, way above me, but I will be checking this thread out.




Is that dish home made or can you get them with with dividers.


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Atheist Chat

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Re: Cloning Pins Directly to Agar [Re: KaptKid] * 1
    #5703665 -

there's supposedly hormones present in your mycelium network that signals to start producing pins. it would then stand to reason that there's a concentration of the hormones left in a very young pin. i still got some problems with it.

i dont think we understand the exact mechanism that causes mycelium to create pins. we understand the conditions required, but that doesn't tell us exactly how it works. maybe i've missed something in my reading but i've never come across anything that conclusively proves there's a hormone involved in the first place, and if so what exactly it is! second off, assuming there is this hormone mechanism, i'd think it would be present in the mycelium network itself. pins are basically "templates" for a full grown mushroom. all of the mushroom's cells are created then and there. they're then inflated with water, and strengthened as they expand. who says the hormone that triggered the pin creation would ever even find its way into the pin then? again there's a hole in my knowledge there as to what the supposed hormone "is"... maybe it has been isolated and pins tested and proven to contain it... so i could be wrong.

its still interesting as hell and i really hope we can all make some conclusions from it.


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Cloning the easy way with LC - newbie friendly pictorial tek

All your answers are here!

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703684 -

^ mysterious...just another reason why I love mushrooms.

RR, as RK said, you seem to make the *seemingly* impossible possible. I have no doubts you will find what you are looking for. Good luck.


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channel your inner Larry David

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Re: Cloning Pins Directly to Agar [Re: demiu5]
    #5703754 -

btw, how much are those 3 chambered dishes?


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

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

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Re: Cloning Pins Directly to Agar [Re: KaptKid]
    #5703785 -

I don't think the pin would have more hormones than a cutout of the same pin. I also think peroxide is horribly overrated for agar work. It does a good job of 86'ing cobweb on a casing layer, but isn't much good in my experience preventing bacteria on agar. I have some oyster and reishi clones that were taken from wild fruits earier in the week that I dipped in peroxide for five minutes before putting on agar and all six dishes are growing bacteria, not mycelium. You can also dip clones into a 5% bleach solution, and get much better results than peroxide for bacteria and mold spores.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703803 -

have you tried a dilute betadine solution?


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[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

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Re: Cloning Pins Directly to Agar [Re: Psychoslut] * 1
    #5703847 -

I use Gentamicin Sulfate. The three section petri dishes are $68 for a box of 500 from www.cenmed.com You need a business license and an account to get them, but well worth the price. The three sections reduce the price by 1/3 because you can do three experiments simultaneously in the same dish. They're especially practical for strain isolation which uses a lot of agar and dishes. Here's the above dish from five days later.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703861 -

wow !

so it directly grew over the entire pin surface on the agar,
wicked man ! :laugh:
myc looks very healthy and r hizo too


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

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

Edited by ohmatic (06/02/06 12:07 PM)

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703875 -

RogerRabbit said:
I use Gentamicin Sulfate. The three section petri dishes are $68 for a box of 500 from www.cenmed.com You need a business license and an account to get them, but well worth the price. The three sections reduce the price by 1/3 because you can do three experiments simultaneously in the same dish. They're especially practical for strain isolation which uses a lot of agar and dishes. Here's the above dish from five days later.
RR




Good work! That is great. But cubes pin really well anyways so how could you possibly tell if the hormones or whatever you was talking about are actually making a difference?


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[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

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Re: Cloning Pins Directly to Agar [Re: Psychoslut]
    #5703893 -

we shall find out if the crop originating from a pin or from a cloned mature fruit is actually the better performer .
in this case.

right ?


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

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

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Re: Cloning Pins Directly to Agar [Re: RogerRabbit]
    #5703969 -

so you were holding out on us and already knew the odds before asking? :wink:

good exercise in deductive reasoning with instant gratification instead of making us wait that 5 days.  :thumbup: :smile:


--------------------
Cloning the easy way with LC - newbie friendly pictorial tek

All your answers are here!

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Re: Cloning Pins Directly to Agar [Re: ohmatic]
    #5704015 -

ohmatic said:
we shall find out if the crop originating from a pin or from a cloned mature fruit is actually the better performer .
in this case.

right ?


What I'm saying is that even if he does get some incredible flushes from these clones it still wont prove that pin clones perform better because of those hormones. You can get incredible flushes and performance from multi spore inoculations too.


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



[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

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Re: Cloning Pins Directly to Agar [Re: Psychoslut]
    #5704017 -

awesome!


--------------------
Sorry, my english is really bad!

If you like my work, rate me....

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Re: Cloning Pins Directly to Agar [Re: Psychoslut]
    #5704032 -

Psychoslut said:
You can get incredible flushes and performance from multi spore inoculations too.


i totally agree on that.


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

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

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Re: Cloning Pins Directly to Agar [Re: ohmatic]
    #5704046 -

well there is a way to test it.

start with a cloned/isolated strain.  grow it out.  clone a pin and clone a piece of tissue from a mature fruit body.  grow it out again, (2 trays now, same genes, one from pin one from fruit) observe any differences.  repeat experiment from the beginning a few times to eliminate "noise" from your grow parameters and other variance, and see if the ones from the pins consistently do any better.  this way you're dealing with the same genetics all around, but can get back to the original issue, does cloning from a pin have any benefit from cloning at other stages.

i'd bet RR is already ahead of us and has something like that in mind :smile:  just wanted to point it out for the rest.


--------------------
Cloning the easy way with LC - newbie friendly pictorial tek

All your answers are here!

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