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Nan (Nanook)
Posted on Monday, December 10, 2001 - 09:45 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

From a recent series of fruitings on various substrates I am going to report an impression about Cloning. This is not law, just an eye opinion and several months experience working with several "master" clone solutions.

Clones appear to be Substrate specific. For example, if you clone an exceptional fruit growing on cased straw, you will not get the same shroom if fruiting the clone on PF cakes or cased rice... Spawn straw again with the clone... There it is. Clones only run true if you keep the substrates identical (or close to) the initial Tek... In fact I should say clones are TEK SPECIFIC.

Oh, your clone mycelium will fruit on just about any substrate... But clone a giant growing on a PF cake and spawn straw... It's not the same shroom.

CLONE THE LARGEST FRUIT GROWING IN A CLUSTER

Sometimes if you change Teks... The cloned shroom will not appear: but the PF Cake, Invitro Jar, Straw Bed, or Cased Bed will always produce nicely if you clone from a cluster of fruits. Cloning large solitary shrooms may lead to reduced yields if you change Teks and the clone is not happy. A healthy clone will fruit on any proven substrate, but appearance, size, and weight, to me, look very Tek subjective.

Cloning & Dextrose

My clones are carrying forward in pure dextrose and my inoculum continues to run hot and true.

The Tek for carrying cloned strains forward is to limit the number of generations in liquid culture (at least this works for Dextrose). Make a master culture in 100cc's 4% Dextrose solution from the original cloned mycelia. You want a short, ideal incubation, then fridge ASAP. Shoot enough of the culture to be sure it's what you want.

Then use your master to shoot 100cc Dextrose jars. I barely let the master culture warm up: I pull it out of the fridge, Sanitize it, open it in the Glovebox, draw a cc or two out, and shoot another Dextrose jar.

The fresh dextrose jar is Incubated for a few days (really no more than 5-7 days, usually 3-5) and shot into a jar of solid substrate, grown out and fruited.

As your Tek changes, go back to Spores and find a better shroom. But once you find the right shroom, the clone is sustainable for months and months. Work from a master culture, a few cc's at a time, using tight sanitary procedures. You can produce several thousand cc's of cloned inoculum without difficulty.
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onediadem (Onediadem)
Posted on Monday, December 10, 2001 - 11:08 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Sweet.
Nan, your great. I cant wait to be an old pro at this, and be able to look back and laugh.
pj
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jared (Jared112)
Posted on Monday, December 10, 2001 - 02:44 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Good observation.
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hippie3 (Hippie)
Posted on Monday, December 10, 2001 - 03:08 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

good observations.
it makes sense that a clone will do best when fruited on the same substrate, what we are cloning is the genetic expression which did so well on a specific substrate.
change the substrate, and that particular genetic expression will no longer be in it's optimum environment.
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Cragith Kilbonith (Kilborn)
Posted on Monday, December 10, 2001 - 03:54 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

thanks nan, so if you cloned the biggest mushroom from the 9ner tek would it grow better on the 9ner tek or on pints?
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hippie3 (Hippie)
Posted on Monday, December 10, 2001 - 05:00 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

that's an 'interesting' question seeing as how one can use pints with 9er tek.
what are you trying to get at ?
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An guy (Boomer)
Posted on Monday, December 10, 2001 - 08:21 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Regarding this in general, not cloning per se, but the aptness for a fruit for a given substrate, I made sure I printed the largest and the smallest in my prints off a cake. Theory being the smaller ones might express better on a different type of medium- sort of like my own little version of keeping the biodiversity.
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UnkyHerb (Notapplicable)
Posted on Monday, December 10, 2001 - 08:37 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

I've been wanting to clone a few monsters.. but all I have is my oven and I feel it's not as sterile as it should be. So, in fear of failure I havn't attempted to clone anything. Nan, is it hard to successfully do it? And how do you think I should go about doing it with just an oven? Invest in a glovebox would be a good idea wouldn't it.. hehe..

And one more question, on the shroomery, there is a cloning tek involving taking a piece of the mushroom(stem), grinding it in a coffee grinder with some water, then adding h2o2 and letting it sizzle for a bit, then sucking it up and using it to shoot jars. Anyone ever tried that?

And PLEASE, no links to the archives, i've read them over and over ;) Nan, I love what you did.. now you have little helpers posting links directly to the archives, brilliant idea.
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Cragith Kilbonith (Kilborn)
Posted on Monday, December 10, 2001 - 09:32 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

im wondering if you clone the mushroom from the 9ner tek and then you birthed some pf tek and others 9ner tek which would turn out better if cloned?
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Kaijan (Kaijan)
Posted on Monday, December 10, 2001 - 09:42 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Didn't Nan just say the Tek in which the clone originated would best support the clone's fruiting?
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Karna (Karna)
Posted on Monday, December 10, 2001 - 11:03 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Nan, I seek a clarification
When you say that if you clone a bigun from say straw and then use that to spawn say rye, it's not the same shroom, you mean their appearance is different, correct?
Do you think that maybe these new shrooms are identical in other respects, say speed of fruiting, potency, etc except for their appearance? Consequently, when spawned to the same substrate, will the appearance be exactly the same?

Here's why I ask :
I have used this method to clone recently. I spawn a straw bed, pick a really big busty beauty(!) and clone it. I must point out that these are usually singles, not clusters. My straw doesn't really flush, it just pushes out a couple every now and then for weeks. I really don't have the space to do much with straw consistently, so I spawn it to rice or brf which is used to spawn a bulk rice substrate which is fruited in pans. This has resulted in GUT BUSTING first and second flushes. As you say, the appearance is not the same as it was on straw, but I think that is to be expected. In other respects, though, I do think that the clones preserve the characteristics of the source insofar as they are remarkable fruiters.
I must add that this has been done only with PF regular strain. I recently cloned a gorgeous Indian seductress, but that is still in the works. Further, I have never, unfortunately spawned the straw clone to more straw. Maybe next time.

So what do you say, you think that clones are identical in everything except appearance, or you think the substrate changes everything?
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Nan (Nanook)
Posted on Tuesday, December 11, 2001 - 12:24 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Appearance and growth characteristics both seem to change as you change the environment. With two widely diffeent Teks you end up with shrooms that look completely, completely different. The further you go from the clone's "ideal" substrate, the smaller the stature, the less hefty the yield, the less resistant to contams (I include contam resistance in my evaluation of clones)... Looking at them it simply blows my mind how different the fruit are.

It struck me after I provided some clone culture to a friend who used all different Teks in an all different environment... And his shrooms didn't look _anything_ like mine and there was _no_ chance of a mixup... And I have seen this happen before, because I have evaluated clones in different Teks.

Good hunting!

P.S. A cardboard glovebox is better than an oven. The only thing I trust doing over an oven is shooting PF jars, but you can shoot PF jars 100% contam free in a decent still, clean, room.

There is no way in hell I would attempt to clone or work with liquid cultures in a oven. Maybe I am narrow minded, I find the oven awkward and inconvenient, hot, and my sterile technique suffers. Give me a clean comfortable box any day, even a well disinfected cardboard box with some playtex gloves... There is an element of confidence and the ability to work at a smooth steady pace. I take off the gloves and I _know_ everything was sterile. That's nice.
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Cragith Kilbonith (Kilborn)
Posted on Tuesday, December 11, 2001 - 12:35 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Kaijan i understood it when i read it, wondering cause they are basically the same thing, its not straw or anything. i was just wonderin
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hippie3 (Hippie)
Posted on Tuesday, December 11, 2001 - 01:30 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

they are both essentially the same--rice.
one is ground, the other left whole.
so the cloned specimen from one should work fairly well on the other.
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Cragith Kilbonith (Kilborn)
Posted on Tuesday, December 11, 2001 - 06:27 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

thanks hippie. thats all i was wondering, ill test it out one day and report my final results (dont count on it any time soon ).
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Karna (Karna)
Posted on Thursday, December 13, 2001 - 10:51 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Something to report for stats sake

A foaf recently cloned a beautiful specimen from straw and inoculated two pints of pf substrate with this. Nan, this is the method I described in your cloning thread. One pint spawned a straw tray. This, unfortunately was accidentally left sealed for a week while she was on vacation and succumbed.

The second spawned 1 regular size bag of Lundberg brown basmati rice using a bulk bag tek (1 bag, I think it's either 1 or 2lb rice, yields about 9 pints cooked substrate). The bag was colonized more or less completely in 7 days. At this point it was cased with 1/2 peat, 1/2 verm. Lime was omitted by accident.

The flush from this pan was by far the most stunning display she, and myself had ever seen. When viewed from the top, nothing was visible of the pan itself or any stems. It was basically a canopy of caps. Not a single abort.

The yield when measured with a hanging postal scale (not known for hair splitting accuracy) was altogether about 135 gms dried. This I figured cannot be right, so a digital is going to be pressed into service when she can find one. Whatever the error, I don't suppose it is more than maybe a dozen gms on either side. In any case, that still makes it a bumper freaking crop.

2 downers
Firstly, the cloned strain was lost since by the time she decided to make a jar of clone water, the master clone solution had perished. Also she did not want to reclone any of the fruit. So it's back to another straw tray in the works. She is certain however that the results are quite easily replicable. Which brings me to downer 2-
At this time, pic capability is STILL not on although hints have been dropped for Christmas, so we'll see.

Finally as a nod to Nan's cloning thread, the fruit looked NOTHING like the cloned straw beauty. However, they grew quick, fruited very fast and VERY impressively, like their patriarch.

I'll repost yield when the digi is located, hopefully (if urge to consume is resisted) along with number of fruit bodies.
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hippie3 (Hippie)
Posted on Friday, December 14, 2001 - 04:22 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

sounds pretty impressive.
what strain was it ?
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Karna (Karna)
Posted on Friday, December 14, 2001 - 10:39 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

pf regular strain- my #1.
Ok Digi was pressed into service and my above result was off by 14gms for a total of 121gms.
Sorry I realized I just do not have the patience to count all those little fuckers - a couple hundred or so?