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OfflineGrizzdude
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isolating vs. cloning
    #12036673 - 02/15/10 08:54 PM (14 years, 10 months ago)

what is the difference between isolating and cloning?


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Offlineoh_you_know
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Re: isolating vs. cloning [Re: Grizzdude]
    #12036684 - 02/15/10 08:57 PM (14 years, 10 months ago)

good question. from my understanding, cloning is almost an exact genetic copy of the fruit you clone. isolating is getting a few strains then fruiting them to see which one you like best then sticking with it. i assume once you iso, you either keep using the iso or clone your best fruit from the iso. would a few generations of clones be considered an iso if you kept taking the largest fruit?


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Offlinecookeman
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Re: isolating vs. cloning [Re: Grizzdude]
    #12036735 - 02/15/10 09:09 PM (14 years, 10 months ago)

Isolating is typically done to single out one strain from many. For example: when you have a spore print there are many many many different strains. In order to single out one really good strain you can swab an agar petri dish with some of the spores from the print. After a while the spores will begin to germinate and you'll still have many many strains to choose from. You then take wedges from the best looking strains and start new petri dishes, and continue to do so until you have one single strain called an "isolate".

A clone can be made several different ways. Whether it be from a small pin, or a section of mature mushroom or really anything you're making a duplicate of. You can make a clone of an isolate because you're just duplicating the same single strain. There's no need to isolate a clone because it's already an isolate aka single strain.

Hope that clears things up


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OfflineGrizzdude
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Re: isolating vs. cloning [Re: cookeman]
    #12036896 - 02/15/10 09:38 PM (14 years, 10 months ago)

ok so I guess i want to "clone" this very potent strain I found to keep it going?


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OfflineMOPE
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Re: isolating vs. cloning [Re: cookeman]
    #12037050 - 02/15/10 10:05 PM (14 years, 10 months ago)

Quote:

cookeman said:
Isolating is typically done to single out one strain from many. For example: when you have a spore print there are many many many different strains. In order to single out one really good strain you can swab an agar petri dish with some of the spores from the print. After a while the spores will begin to germinate and you'll still have many many strains to choose from. You then take wedges from the best looking strains and start new petri dishes, and continue to do so until you have one single strain called an "isolate".

A clone can be made several different ways. Whether it be from a small pin, or a section of mature mushroom or really anything you're making a duplicate of. You can make a clone of an isolate because you're just duplicating the same single strain. There's no need to isolate a clone because it's already an isolate aka single strain.

Hope that clears things up




I thought a clone (not a clone of an isolate) contained a small amount of similar strains? what source said that a clone is always a single set of genetics?


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OfflineMycjunky
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Re: isolating vs. cloning [Re: Grizzdude]
    #12037188 - 02/15/10 10:28 PM (14 years, 10 months ago)

When you clone a certain mushroom it can often contain multiple strains. I believe an isolate would be 1 single substrain that has been totally isolated, typically using agar so you can be sure it's a true isolate of just 1 single strain.

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OfflineMOPE
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Re: isolating vs. cloning [Re: Mycjunky]
    #12037236 - 02/15/10 10:37 PM (14 years, 10 months ago)

Quote:

Mycjunky said:
When you clone a certain mushroom it can often contain multiple strains. I believe an isolate would be 1 single substrain that has been totally isolated, typically using agar so you can be sure it's a true isolate of just 1 single strain.




so a strain can contain multiple sets of genetics, but a substrain has only one set of genetics and is considered an isolate?

sorry if im getting it wrong ha


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OfflineGrizzdude
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Re: isolating vs. cloning [Re: MOPE]
    #12037358 - 02/15/10 11:06 PM (14 years, 10 months ago)

So how do I go about saving this potent strain I have fruiting right now?


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OfflineMycjunky
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Re: isolating vs. cloning [Re: MOPE]
    #12037362 - 02/15/10 11:07 PM (14 years, 10 months ago)

Quote:

MOPE said:
Quote:

Mycjunky said:
When you clone a certain mushroom it can often contain multiple strains. I believe an isolate would be 1 single substrain that has been totally isolated, typically using agar so you can be sure it's a true isolate of just 1 single strain.




so a strain can contain multiple sets of genetics, but a substrain has only one set of genetics and is considered an isolate?

sorry if im getting it wrong ha




Exactly that is how i understand it.

As far as how to save your strain kinda depends what your capable of. If you can do agar work do agar work cause that's typically the most successful way to do it. However you could just clone the mushroom from a flush of this "potent strain" and it would be a good enough clone to serve your purposes most likely. I normally do just that because it's rather hard for me to work with agar due to a high mold spore count and I haven't gotten around to buying a flowhood yet.

Edited by Mycjunky (02/15/10 11:10 PM)

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Invisibleshymanta
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Re: isolating vs. cloning [Re: Grizzdude]
    #12037793 - 02/16/10 12:51 AM (14 years, 10 months ago)

Under as sterile conditions as possible, tare the stem in half length-wise.  With a flame sterilized utensil, rip out several small chunks and place them each on their own agar filled petri dish.

OR

Pick a fresh pin and put it into a vacuutainer, or equivalent, with agar and you've got your master clone you can start new grows with.  Let it colonize and put it into the fridge.

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Re: isolating vs. cloning [Re: MOPE]
    #12038114 - 02/16/10 02:19 AM (14 years, 10 months ago)

Quote:

MOPE said:
Quote:

Mycjunky said:
When you clone a certain mushroom it can often contain multiple strains. I believe an isolate would be 1 single substrain that has been totally isolated, typically using agar so you can be sure it's a true isolate of just 1 single strain.




so a strain can contain multiple sets of genetics, but a substrain has only one set of genetics and is considered an isolate?

sorry if im getting it wrong ha




When we speak about Psilocybe cubensis as a mushroom species, many people in here use the term "strain" while they actually mean the name that was given to a mushroom that was found somewhere in the world.
This is actually not correct! A strain is a result of  "breeding" i.e. genetic narrowing process that reproduces the same phenotypic features over and over again.
Since a single spore print of a mushroom species contains millions of genetic individuals that might all differ in their phenotypic features, it can't really be considered a genetically stable "strain".
A single sector isolate, however only contains the genetic information of two individual spores (at least when you grow them from the isolated mycelium).
This isolate would be considered a strain.
The spore print of this strain will contain millions of different genetic information again, though.
The genetic diversity can be narrowed down, by keep taking spore prints from the best performing isolates (or the isolates that develop the features you're looking for), that developed from the previously taken spore print. This, however takes many, many runs and does not guarantee, that the phenotypes that eventually develop from the spores will show the characteristics you're looking for. PE is a good example for this. Although it's tendency to develop penis shaped fruits is extremely high, it's capable of developing totally normal looking fruits.
A substrain, on the other hand is something different.
Currently a substrain defines itself by showing taxonomic differences from the actual species, such as a different spore size etc. As far as I know, Psilocybe subcubensis is the only substrain of Psilocybe cubensis that is yet know. So it's probably nothing to really think about when it's about genetics of different "strains".
As said correctly above, with cloning you might get multiple strains, since it seems that fruit tissue can contain more than one strain.

Quote:

So how do I go about saving this potent strain I have fruiting right now?




The best thing would probably be to clone a fruit and isolate the -probably- appearing strains.
Let's say you have three sectors of mycelium growing from that tissue, you'd have to isolate every sector and grow it out to know what strain the potent one was. Once you've found it, you can keep cultivating the strain from your master slant.

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Invisibleanonjon
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Re: isolating vs. cloning [Re: Fahkface]
    #12038579 - 02/16/10 07:41 AM (14 years, 10 months ago)

a clone in many cases IS an isolate. clone tissue doesn't always sector. even if a fruitbody does have a few strains co-operating, they are close enough to form fruitbodies together, they close enough to call an isolate. People are just anal retentive and like to brag about their isolation work, so they discount cloning. 3 or 4 genetic variations is close enuff when you're comparing it to multispore which has millions.


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InvisibleDoc_T
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Re: isolating vs. cloning [Re: anonjon]
    #12038595 - 02/16/10 07:50 AM (14 years, 10 months ago)

Quote:

anonjon said:
even if a fruitbody does have a few strains co-operating, they are close enough to form fruitbodies together, they close enough to call an isolate.




It's bound to be better than multi-spore, right?


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Invisibleanonjon
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Re: isolating vs. cloning [Re: Doc_T]
    #12038610 - 02/16/10 07:57 AM (14 years, 10 months ago)

Quote:

Doc_T said:

It's bound to be better than multi-spore, right?




For sure, but you know someone gonna get indignant and argue with this.


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InvisibleDoc_T
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Re: isolating vs. cloning [Re: anonjon]
    #12038620 - 02/16/10 08:01 AM (14 years, 10 months ago)

I've wondered about taking a tissue clone, growing it out, then cloning one of those fruits.
Seems to me, that's a faster way to get to a better (not necessarily perfect) genetic line.


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Re: isolating vs. cloning [Re: anonjon]
    #12038714 - 02/16/10 08:43 AM (14 years, 10 months ago)

It's pretty dependent on what you want to do with it. If you're planning on creating your personal "ultimate strain", further isolation would be a good idea. If you're cloned tissue produces what you want over and over again anyway, there's certainly no need for further isolation.
Single sector isolation might be necessary for commercial mushroom farms, but that's pretty much it.
If you want to find out of how certain circumstances have an influence on the growth of a mushroom, single sector isolation is just a little more accurate than cloning. It might not even be necessary, depending on how many -if at all- strains are present in the fruit.
Cloning is definitely a pretty easy and rather fast way to get what you want.

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OfflineGrizzdude
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Re: isolating vs. cloning [Re: Fahkface]
    #12038977 - 02/16/10 09:56 AM (14 years, 10 months ago)

Ok so I'll start out by putting some pieces of shroom tissue in a few agar dishes. What carictaristics do I need to look for when the myc starts growing? What is this sectoring that I keep hearing? Will the myc grow differently in the same dish or something, not too sure what that means?


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Re: isolating vs. cloning [Re: Grizzdude]
    #12039625 - 02/16/10 12:16 PM (14 years, 10 months ago)

Quote:

Grizzdude said:
Ok so I'll start out by putting some pieces of shroom tissue in a few agar dishes. What carictaristics do I need to look for when the myc starts growing? What is this sectoring that I keep hearing? Will the myc grow differently in the same dish or something, not too sure what that means?




Here you get an idea of the cloning process. If you see more than one "kind" of mycelium growing, your tissue probably consisted of more than one strain.
Watch this to get an idea what follows next.

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Re: isolating vs. cloning [Re: Fahkface]
    #12039708 - 02/16/10 12:31 PM (14 years, 10 months ago)

IME cloning has always given me 1 isolate. So for me its a lot easier to just clone desired fruits and put them on agar, do another transfer just for fun and then use that dish for work.


Its much more efficient to start with a clone then to start from spores. Plus after getting isolates from spores you still have to grow them to find the one with desired traits. When you do a clone you already pick the fruit you want to clone, so you eliminate that added step.


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OfflineFahkface
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Re: isolating vs. cloning [Re: teesionbear]
    #12039782 - 02/16/10 12:46 PM (14 years, 10 months ago)

By the time a mushroom fruits, plenty of strains have exchanged genetic information while colonizing the substrate. The fruits contain all these information.
The fruits that show up might be good, but also might be far from ideal in terms of what "the genetics actually have to offer".
A spore, on the other hand is a genetic individual.
If the fruit you cloned is exactly what you desired, there's certainly no need to do anything but growing it out over and over again.
If you want to optimize even further, you'd have to start from spores and isolate before the strain-merging has taken place.
This way your chance to discover the best performing strain is a lot higher.

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OfflineGrizzdude
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Re: isolating vs. cloning [Re: Fahkface]
    #12040128 - 02/16/10 01:43 PM (14 years, 10 months ago)

Thanks for the help guys, I'll be checking out those links Fahkface! Thanks!


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