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Offlineforevrgrounded98
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Registered: 10/25/03
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when cloning
    #2072524 - 11/05/03 07:44 AM (20 years, 4 months ago)

Is putting the chunk of the inside of the mush on a petri dish the only way to clone? If it is, are there any everyday types of stores that cary petri dishes filled with the solution? I've never seen them or heard of a place to get them. Thanks
-matt


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"If I don't see ya before the end of this one, I'll meet ya in the next one and don't be late" - Jimi Hendrix

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OfflineRoseM
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Re: when cloning [Re: forevrgrounded98]
    #2072535 - 11/05/03 07:49 AM (20 years, 4 months ago)

Search for the Poor Man's Syringe Tek and the Karo Tek.

These are two other ways to clone. Not as precise as agar but still cloning.

Basically, any time you grow mycelia from a chunk of a mushroom, you are cloning that mushroom.


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Fiddlesticks.


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Invisiblemicro
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Re: when cloning [Re: Rose]
    #2072549 - 11/05/03 07:56 AM (20 years, 4 months ago)

You can use mycelium, too. If you're growing it in bags, you can inject sterile water, mix some mycelium into it, and draw it back with a sterile syringe (using rubbing alcohol, etc.) Just try not to let the water touch the mycelium, or it will absorb it, and you'll be left with very little solution.

This is an easy way, and you don't need a glovebox, or anything, but you really do need to be growing in bags to do this (it's a lot harder to do with jars.)

Basically, you can clone with any live tissue.

--
Micro


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OfflineRoseM
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Re: when cloning [Re: micro]
    #2072554 - 11/05/03 07:59 AM (20 years, 4 months ago)

Micro is right. That is the easiest way to clone, but if you clone your biggest shroom, you should get clones of your biggest shroom. If you clone the mycelia from a multispore inoculation, you'll still get shrooms, but you won't get many of the benefits of cloning.


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OfflineDMJ
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Registered: 10/10/03
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Re: when cloning [Re: micro]
    #2072655 - 11/05/03 08:48 AM (20 years, 4 months ago)

Quote:

micro said:
You can use mycelium, too. If you're growing it in bags, you can inject sterile water, mix some mycelium into it, and draw it back with a sterile syringe (using rubbing alcohol, etc.) Just try not to let the water touch the mycelium, or it will absorb it, and you'll be left with very little solution.



--
Micro





Could you specify this a little more for me Micro? What do you inject the sterile water into? What do you mix the myc with? The sentences seem to indicate injecting the water into the myc, but in the next sentence, you say don't let it touch the water.
In no way am I doubting what you say, just confused by the way it was said.


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OfflineRoseM
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Re: when cloning [Re: DMJ]
    #2072694 - 11/05/03 09:03 AM (20 years, 4 months ago)

Search for the poor man's syringe tek DMJ. This is what micro described.


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OfflineRoseM
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Re: when cloning [Re: Rose]
    #2072718 - 11/05/03 09:13 AM (20 years, 4 months ago)



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OfflineDMJ
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Re: when cloning [Re: Rose]
    #2072760 - 11/05/03 09:38 AM (20 years, 4 months ago)

Thank you Cervantes!


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I come alive in the darkness.

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Offlinethisone
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Re: when cloning [Re: DMJ]
    #2072853 - 11/05/03 10:32 AM (20 years, 4 months ago)

So cloning is any live tissue, and an isolate is different because that is from a specific fruit body, or is that selected from sub strains rhizo's selected and picked apart from an agar plate and fruited for particular characteristics? Sorry, I'm a bit confused on this issue.


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Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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OfflineRoseM
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Re: when cloning [Re: thisone]
    #2072868 - 11/05/03 10:38 AM (20 years, 4 months ago)

An isolate is selected sub strains on agar.

Cloning a specific shroom, is like selecting a good fruiting isolate though. It may not be perfect, but you know it works in your growing environment.

They are both forms of cloning, you can only get a pure strain on agar, as I understand it.


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Invisiblemicro
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Re: when cloning [Re: Rose]
    #2072895 - 11/05/03 10:50 AM (20 years, 4 months ago)

1.) Inject sterile water into bag (sterilly -- use rubbing alcohol.) Don't let it touch the mycelium; just let it fall into the bottom corner, holding the mycelium away from it.

2.) Without opening t/ bag, break off a little piece of mycelium and mix it into the water by smooshing it.

3.) Take a sterile syringe (like the one you used for the water if you kept it "sterile") and draw up your mycelium water.

I don't think cloning the biggest mushroom will change anything; the genetics should be the same throughout the mycelium (the mushrooms are all one organism.)

I've heard a lot of people say that, but it seems like a myth. Bigger mushrooms are easier to work with, though, anyway.

--
Micro


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

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OfflineRoseM
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Re: when cloning [Re: micro]
    #2072977 - 11/05/03 11:23 AM (20 years, 4 months ago)

Hmm, I've often wondered about the biggest shroom, definately cloning from good fruiting mycelium is wise.

If you do a multispore inoculation of several jars, cloning from your best cake is wise.


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Offlinethisone
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Re: when cloning [Re: Rose]
    #2073055 - 11/05/03 11:49 AM (20 years, 4 months ago)

but isn't the purpose of cloning a fruitbody to gurantee that you have a fruiting strain. I would think that the biggest should be selected because then that fruitbody also has the charcteristics of being a large fruiting strain. Taking a sample from the mycelilum wouldn't ensure it was a fruiting strain would it? What are the benefits of taking a cloned fruitbody or isolating the strain on agar?


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Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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OfflineRoseM
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Re: when cloning [Re: thisone]
    #2073117 - 11/05/03 12:08 PM (20 years, 4 months ago)

That is what I have heard, but I've not experimented enough to know for sure.


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Invisiblemicro
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Re: when cloning [Re: Rose]
    #2073249 - 11/05/03 12:54 PM (20 years, 4 months ago)

It probably has a lot more to do with the distribution of hormones to get big mushrooms or small mushrooms. Genetics certainly plays a part, but the genetics in a mycelium should be the average of the distribution of nuclei in the cells -- one place shouldn't be much different than another.

--
Micro


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)

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Offlineforevrgrounded98
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Re: when cloning [Re: micro]
    #2073350 - 11/05/03 01:28 PM (20 years, 4 months ago)

Why wouldn't it work to take a sample of the inside of a stem and instead of mycelium, mash up the inside of the stem and add it with water to suck it up in a syringe? Sorry, if that sounds dumb.
-matt


--------------------
"If I don't see ya before the end of this one, I'll meet ya in the next one and don't be late" - Jimi Hendrix

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OfflineRoseM
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Re: when cloning [Re: micro]
    #2073997 - 11/05/03 04:42 PM (20 years, 4 months ago)

True, but I've wittnessed vastly different looking mycelium from the same spores. If you start from multispore, rather than an isolate, I'd still recomend waiting for fruit, to see which myc is most worth cloning if you start from a multispore innoculation.

I definately see what you mean about the size of the fruit not mattering, they all come from the same myc. I always found that theory a little odd. Hormones make more sense to me. This would be great if you isolated a good strain and grew it on some substrate. You could get your money's worth from the poor mans syringe tek.


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Fiddlesticks.


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OfflineRoseM
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Re: when cloning [Re: forevrgrounded98]
    #2074007 - 11/05/03 04:45 PM (20 years, 4 months ago)

Forever,

You are very close to the correct way to clone a shroom.

If you cut the stem open with a sterile blade, the inside of the stem will be sterile. Take a piece of myc from the inside with your blade and use it with whatever medium you need to grow or inject the mycelia.


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Fiddlesticks.


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Offlineforevrgrounded98
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Re: when cloning [Re: Rose]
    #2074213 - 11/05/03 05:40 PM (20 years, 4 months ago)

ok, so let me see if i can dumb things down a shade for myself....In a sterile environment, if i took a properly prepared substrate and innoculated it with that sterile bit of mush you were talking about, would it colonize the substrate like an innoculation shot would?


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"If I don't see ya before the end of this one, I'll meet ya in the next one and don't be late" - Jimi Hendrix

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OfflineRoseM
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Re: when cloning [Re: forevrgrounded98]
    #2074257 - 11/05/03 05:53 PM (20 years, 4 months ago)

It could... as long as the piece of shroom did not dry out (I learned the hard way... dry=dead). You could soak the piece in h2o2 to hydrate and sterilize it... if that is still simple enough for you :wink:.


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Fiddlesticks.


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Offlineforevrgrounded98
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Re: when cloning [Re: Rose]
    #2074285 - 11/05/03 06:01 PM (20 years, 4 months ago)

aw yea, nice and dumb, just like i like it :smile:  So is there a certain dilution of h202 i should use or just let it sit in a cup of h202 right from the bottle for a few hrs?


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"If I don't see ya before the end of this one, I'll meet ya in the next one and don't be late" - Jimi Hendrix

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Offlinethisone
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Re: when cloning [Re: forevrgrounded98]
    #2074461 - 11/05/03 06:59 PM (20 years, 4 months ago)

I can ssee the benefit of growing on agar to find an isolate strain specifically focused on the rhizomorphic are which is more likely to fruit. A cloned fruit body is guaranteed to be a fruiting strain since it has already been tested in that current grow. hormones sounds right to me too although I have seen projects with cloned fruitbodies where they were first grown on cakes using a multi-spore innoc the grown again under the same parameters with a much higher yeild low abhort rate and the fruitbodies that grew were all relatively similar in size. What would account for this? Does anyone know the benefit of growing an isolate out on agar compared with cloning a single fruitbody?


--------------------
Everything written above this line is completely false. I am terribly bored and write here to pass time. I do not participate or support any illegal activity.

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OfflineRoseM
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Re: when cloning [Re: thisone]
    #2075236 - 11/05/03 10:03 PM (20 years, 4 months ago)

I'd try that first forevr... but 10/1 Water/h2o2 is a common measurement.

I realy doubt pure h2o2 would harm the mycelia enough to kill it all.


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InvisibleShamanorrhea
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Re: when cloning [Re: Rose]
    #2075852 - 11/06/03 12:59 AM (20 years, 4 months ago)

Cervantes, pure peroxide can be detrimental to mycelial growth. However, the mycelia will usually recover and begin to flourish after the initial shock of being exposed to peroxide.

There is a link on this site for a cloning procedure. I'm not too sure if it is useful or not but I'll post it anyways.

http://mv.lycaeum.org/mu/_liquid_culture.html

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OfflineRoseM
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Re: when cloning [Re: Shamanorrhea]
    #2075888 - 11/06/03 01:11 AM (20 years, 4 months ago)

Yeah I know that, but if you are taking a piece of shroom, I think the myc has a real fighting chance. I always hear h2o2 is detrimental to growth bu,t I've never heard it kills mycelia outright. It will stun the myc and a 10/1 water ratio... or even 2/1 would be better... but since we're talking about the lazy way, I think pure h2o2 is the simplest way to insure sterility and hydration of the mycelia.

I'm not saying it would be the first thing I'd try... I just like lazy tek theory. :smile: The more h2o2 you can use... the less need for a flowhood. A lazy person has no use for a high tek contraption like a flowhood or a glovebox! Those take work!

Cool link.



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OfflineTrippinSpinners
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Re: when cloning [Re: forevrgrounded98]
    #2075893 - 11/06/03 01:13 AM (20 years, 4 months ago)



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OfflineRoseM
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Re: when cloning [Re: TrippinSpinners]
    #2075896 - 11/06/03 01:14 AM (20 years, 4 months ago)

Looks fun.


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