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InvisibleWolfgang
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cloning and strain isolation
    #7895760 - 01/18/08 12:54 AM (16 years, 15 days ago)

I've researched this for the past couple days and still have few questions regarding cloning and strain isolation. When mushroom mycelium sectors it's really just the different strains right? If we do a enough agar transfers we eventually end up with ONE strain.

If we were to take a clone from this strain would it still sector? How do you get the best of both worlds an isolated strain that performs great but is also a clone? Do I just a clone from an isolated strain of mycelium? and if I were to clone an aborts would I get aborts? sry for all the questions


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Invisiblezyxwvutsr
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Re: cloning and strain isolation [Re: Wolfgang] * 2
    #7895838 - 01/18/08 01:23 AM (16 years, 15 days ago)

No. When agar is inoculated and more then one sector of mycelium grows, these are sub-strains.

Isolating a sub-strain is what you are wanting to do. I personally took a print knocked up some agar, isolated the most rhizo myc (after trial and error, I ran into a non fruiting and low active sub-s.) and made an LC. LC to WBS, WBS to Poo, took print from nicest mush, and started process over.

This would be isolating a substrain and then cloning it. I have used same strain for four years in a row. All from one LC containing cloned sub-strain


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Invisiblezyxwvutsr
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Re: cloning and strain isolation [Re: Wolfgang]
    #7895855 - 01/18/08 01:28 AM (16 years, 15 days ago)

"do I just clone from an isloate sector?"

Yes. Isolate your wanted sub-strain. Grow your mushrooms. take a print from a healthy strong mushroom, start over. If everything is right, when agar is knocked up with your chosen mushrooms spores, you swhould have one section of growth.

I don't know about aborts, But I have cloned short and stubby to defected/mishapen mushes(not due non-optimal parameters) to find growing the same mushroom again and again


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: zyxwvutsr]
    #7895915 - 01/18/08 01:55 AM (16 years, 15 days ago)

At which point do I clone? Why do I need to start from spores again if I've already isolated my sub-strain? I thought spores had many many sub-strains you're saying that once I isolate and fruit a sub-strain it's spores won't sector. So why don't vendors offer spores that don't sector and come from the good sub-strains?


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Invisiblezyxwvutsr
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Re: cloning and strain isolation [Re: Wolfgang] * 1
    #7895941 - 01/18/08 02:10 AM (16 years, 14 days ago)

Once you have isolated a sector, that is a substrain. Grow this substrain. Take a print from a good mush (whichever mush you want to clone) and start over. All spores from here on out are the same thing, over and over.

You could isolate one sector, grow it and be good. But there are many variables uncontrolled.

Vendors offer a multispore syringe. This syringe contains spores all of one strain. They would cost more because it would take more time to prepare, thus reducing profit margins.

Check threads at bottom of this page, sometimes yields betters results then search function. *out for the night*


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OfflineSamurai_Jim
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Re: cloning and strain isolation [Re: zyxwvutsr]
    #7896117 - 01/18/08 05:35 AM (16 years, 14 days ago)

If you get the book The Mushroom Cultivator, there is an entire section dedicated to this specific subject. I learned all I needed to know about the process from there. THen I had to actually try it. Get the book if you don't have it, you will not be disappointed.


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: Samurai_Jim]
    #7909489 - 01/21/08 12:47 PM (16 years, 11 days ago)

bump


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OfflineRogerRabbitM
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Re: cloning and strain isolation [Re: Wolfgang]
    #7909760 - 01/21/08 02:03 PM (16 years, 11 days ago)

Quote:

Yes. Isolate your wanted sub-strain. Grow your mushrooms. take a print from a healthy strong mushroom, start over. If everything is right, when agar is knocked up with your chosen mushrooms spores, you swhould have one section of growth.





Actually, that's not right at all. When spores are used, many strains and isolates will develop. In other words, if you fruit an isolated strain, the spores that fall, will NOT be an isolated strain.

I've written pages and pages on how to isolate strains. Search should reveal all you want to know. There's also a strain isolation tek on my dvd if you learn easier by seeing it done.
RR


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7909960 - 01/21/08 02:47 PM (16 years, 11 days ago)

Quote:

RogerRabbit said:
Quote:

Yes. Isolate your wanted sub-strain. Grow your mushrooms. take a print from a healthy strong mushroom, start over. If everything is right, when agar is knocked up with your chosen mushrooms spores, you swhould have one section of growth.





Actually, that's not right at all. When spores are used, many strains and isolates will develop. In other words, if you fruit an isolated strain, the spores that fall, will NOT be an isolated strain.

I've written pages and pages on how to isolate strains. Search should reveal all you want to know. There's also a strain isolation tek on my dvd if you learn easier by seeing it done.
RR




That's what I thought RR and right now I'm searching through your posts on strain isolation and cloning, I already own both copies of you dvds I just want to make sure my understanding of how get an isolated strain that's also a clone are correct.


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Invisiblesimplemachine
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7910130 - 01/21/08 03:37 PM (16 years, 11 days ago)

I'm still unclear as to the difference between cloning a fruit from an isolated strain, and cloning a fruit from multispore.

Would a clone of a fruit from a multispore inoculation be a clone of a single substrain?


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Invisiblesimplemachine
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Re: cloning and strain isolation [Re: simplemachine]
    #7910241 - 01/21/08 04:07 PM (16 years, 11 days ago)

Found my answer here.

Search feature FTW.


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: simplemachine]
    #7910412 - 01/21/08 04:31 PM (16 years, 11 days ago)

:thumbup:


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OfflineRogerRabbitM
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Re: cloning and strain isolation [Re: Wolfgang] * 1
    #7911018 - 01/21/08 06:21 PM (16 years, 11 days ago)

If you grow a mushroom from an isolated strain, the fruits themselves will be of that isolate, but you wouldn't want to clone from them. You'd want to go to your refrigerator and pull out a much younger specimen of the same mycelium.
RR


--------------------
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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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Invisiblesimplemachine
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7911161 - 01/21/08 06:40 PM (16 years, 11 days ago)

Is that to avoid senescence? Would that mean that their is a wider selection of genes in the original isolate than the fruit?


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OfflineMYSTIQUE
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Re: cloning and strain isolation [Re: simplemachine]
    #7911287 - 01/21/08 06:56 PM (16 years, 11 days ago)

Quote:

simplemachine said:
Is that to avoid senescence? Would that mean that their is a wider selection of genes in the original isolate than the fruit?




The orginal would be younger and stronger faster to colonize and all that.


--------------------
Dont know what the fuck I just said? READ THIS
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I ain't a hippy but I'm covered in dirt                         
Sippin lots of mushroom tea in a tye-dye shirt
Chasin' the Grateful Dead, no shoes on my feet
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InvisibleWolfgang
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7911312 - 01/21/08 06:59 PM (16 years, 11 days ago)

What do we clone from then? How do you get a strain that will have consistently perform well and have good potency?


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OfflineMYSTIQUE
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Re: cloning and strain isolation [Re: Wolfgang] * 1
    #7911361 - 01/21/08 07:05 PM (16 years, 11 days ago)

You have your strain isolated in a slant or dish. You should have a few different strains isolated from the original.

You take a small piece and put it in grain or pf and then you let it fruit.

You see if it has the quality's that you like.

You should have a large slant or dish that you labeled and put in the fridge that goes with your fav jar.

That is now your master culture. Take a small part of it and save the rest.

Take the small part and put it in a new slant or dish and use that to make spawn and what not.


--------------------
Dont know what the fuck I just said? READ THIS
http://www.shroomery.org/5122/The-Shroomery-Mushroom-Glossary
I ain't a hippy but I'm covered in dirt                         
Sippin lots of mushroom tea in a tye-dye shirt
Chasin' the Grateful Dead, no shoes on my feet
Beggin' in the parking lot for something to eat,

:onfire:DO NOT USE FIRE IN YOUR GLOVE BOX!!!!!!!:onfire:


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: MYSTIQUE]
    #7911410 - 01/21/08 07:12 PM (16 years, 11 days ago)

I'm apologize I'm missing something here, I didn't see anything in that about cloning isn't that the only way to have reliable potency?


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OfflineMYSTIQUE
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Re: cloning and strain isolation [Re: Wolfgang]
    #7911599 - 01/21/08 07:43 PM (16 years, 11 days ago)

You are cloning when you cut the section out of the slant and droping it into the jar in a really loose sense of the word. You test out the potency of whats in the jar when its fruited and anything else taken from the isolated strain in the slant that you took the section from will be the same as the fruit you have.


--------------------
Dont know what the fuck I just said? READ THIS
http://www.shroomery.org/5122/The-Shroomery-Mushroom-Glossary
I ain't a hippy but I'm covered in dirt                         
Sippin lots of mushroom tea in a tye-dye shirt
Chasin' the Grateful Dead, no shoes on my feet
Beggin' in the parking lot for something to eat,

:onfire:DO NOT USE FIRE IN YOUR GLOVE BOX!!!!!!!:onfire:


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: MYSTIQUE]
    #7913776 - 01/22/08 05:23 AM (16 years, 10 days ago)

Yeah but I thought potency could vary mushroom to mushroom in any given flush unless you cloned the actually mushroom tissue.

I'm pretty sure I see what you're saying tho.


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OfflineRogerRabbitM
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Re: cloning and strain isolation [Re: MYSTIQUE]
    #7913937 - 01/22/08 09:30 AM (16 years, 10 days ago)

Quote:

MYSTIQUE said:
You have your strain isolated in a slant or dish. You should have a few different strains isolated from the original.

You take a small piece and put it in grain or pf and then you let it fruit.

You see if it has the quality's that you like.

You should have a large slant or dish that you labeled and put in the fridge that goes with your fav jar.

That is now your master culture. Take a small part of it and save the rest.

Take the small part and put it in a new slant or dish and use that to make spawn and what not.




:thumbup:  Couldn't have said it better.
RR


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"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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InvisibleWolfgang
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7913965 - 01/22/08 09:42 AM (16 years, 10 days ago)

I guess my understanding is still pretty flawed then I could have swore I read that potency varies from mushroom to mushroom on any given flush.. including flushes from isolated strains

but I see that can't be true.. thanks


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OfflineMYSTIQUE
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Re: cloning and strain isolation [Re: Wolfgang]
    #7914007 - 01/22/08 10:02 AM (16 years, 10 days ago)

Well potency isn't totally controlled by genetics. Get the best isolate ever and grow it on white rice flour and it will suck.

And thanks RR I learned everything about cloning mushrooms from you.


--------------------
Dont know what the fuck I just said? READ THIS
http://www.shroomery.org/5122/The-Shroomery-Mushroom-Glossary
I ain't a hippy but I'm covered in dirt                         
Sippin lots of mushroom tea in a tye-dye shirt
Chasin' the Grateful Dead, no shoes on my feet
Beggin' in the parking lot for something to eat,

:onfire:DO NOT USE FIRE IN YOUR GLOVE BOX!!!!!!!:onfire:


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: MYSTIQUE]
    #7914397 - 01/22/08 11:58 AM (16 years, 10 days ago)

It's nearly exclusively controlled by genetics according to RR he's grown on everything with no difference in potency between in substrates.


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OfflineMYSTIQUE
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Re: cloning and strain isolation [Re: Wolfgang]
    #7914415 - 01/22/08 12:05 PM (16 years, 10 days ago)

naw you cant use sub to make super strong shrooms but you can use it to make weak ones. Just look up any of the white rice flour grows.


--------------------
Dont know what the fuck I just said? READ THIS
http://www.shroomery.org/5122/The-Shroomery-Mushroom-Glossary
I ain't a hippy but I'm covered in dirt                         
Sippin lots of mushroom tea in a tye-dye shirt
Chasin' the Grateful Dead, no shoes on my feet
Beggin' in the parking lot for something to eat,

:onfire:DO NOT USE FIRE IN YOUR GLOVE BOX!!!!!!!:onfire:


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: MYSTIQUE]
    #7914449 - 01/22/08 12:19 PM (16 years, 10 days ago)

Quote:

MYSTIQUE said:
You are cloning when you cut the section out of the slant and droping it into the jar in a really loose sense of the word.




What's the traditional direct mushroom clone used for then? Wouldn't this definition of a clone differ greatly from that of a cloned mushroom?

When we cut the section out of the slant and drop it in the jar to test for the qualities we like, that flush will have many mushrooms of DIFFERENT potencies correct but if we cloned a specific mushroom that flush would not have different potencies from mushroom to mushroom?


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OfflineRogerRabbitM
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Re: cloning and strain isolation [Re: Wolfgang]
    #7914680 - 01/22/08 01:24 PM (16 years, 10 days ago)

When you transfer from a slant or petri dish, you're not cloning. Cloning is taking live tissue from a fruit. Transferring from a slant or petri is simply transferring.

About the white rice thing. It is a crappy substrate but doesn't produce non-active mushrooms. Many new growers try different things, and thus when they get a bum batch, they often think it was the substrate. Ditto for when they get a killer potency flush. Over the years, I've heard dozens of claims of inactive mushrooms, and extremely potent mushrooms over just about every strain and every known substrate.
RR


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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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InvisibleWolfgang
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7917024 - 01/22/08 08:40 PM (16 years, 10 days ago)

So when do we clone!?


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Invisiblesimplemachine
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7917816 - 01/22/08 10:40 PM (16 years, 10 days ago)

Quote:

So when do we clone!?




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Offlinelaten
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Re: cloning and strain isolation [Re: simplemachine]
    #7917864 - 01/22/08 10:51 PM (16 years, 10 days ago)

You clone from the upper part of the mushroom stem when its done fruiting. Or you could put a pin on a plate and clone that.


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Invisiblethedefone
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Re: cloning and strain isolation [Re: simplemachine] * 1
    #7917948 - 01/22/08 11:08 PM (16 years, 10 days ago)

Here is a good link on sectoring. I'm posting this for my benefit, and for the other idiots like me, who didn't quite know. Also, thanks simplemachine for that link.

My question. At what point does senescence start to kick in? If it takes multiple sectorings to finally establish the ideal strain, how close are you to genetic breakdown at that point? Or am I totally missing it?


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OfflineRogerRabbitM
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Re: cloning and strain isolation [Re: thedefone]
    #7918014 - 01/22/08 11:20 PM (16 years, 10 days ago)

Quote:

how close are you to genetic breakdown at that point?




You're still a long way off from senescence. Mycelium will generally grow aggressively for several months. Your isolations will be done within a month to six weeks. At that point, save all your strains in the refrigerator to stop growth(more accurately-slow the growth way down). When the cells stop dividing, the road to senescence gets much longer. This means that two years from now, you can pull a master slant out of the refrigerator and it will realistically only have that original six weeks worth of cell divisions on it.
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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Invisiblethedefone
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7918082 - 01/22/08 11:34 PM (16 years, 10 days ago)

Okay. So, that will only start being an issue if you are taking clones from, say, isolated fourth-flush fruit bodies.

Also, when you take sectors and transfer them to new plates, they do continue to re-sector themselves, no? But, the genetic variation within those resulting sectors is narrower, yes? Just thinking out loud...


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InvisibleWolfgang
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Re: cloning and strain isolation [Re: RogerRabbit]
    #7918825 - 01/23/08 05:56 AM (16 years, 9 days ago)

Would a proper agar isolate be considered more "pure" in a genetic sense than a culture from a cloned tissue sample that was taken from an isolated line of mycelium?

Would a cloned tissue sample taken from an isolated line of mycelium PERFORM like it's mycelium would?

How can a cultivator have consistently great results in terms of potency AND performance?


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Invisiblethedefone
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Re: cloning and strain isolation [Re: Wolfgang]
    #7926804 - 01/24/08 09:21 PM (16 years, 8 days ago)

bump.


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Re: cloning and strain isolation [Re: thedefone]
    #8016395 - 02/13/08 06:22 AM (15 years, 11 months ago)

yeah i was getting bad potency on straw pf tek then i got some poo
from a sponsor grew out albinos and before i was eating maybe 300 grams of wet from my pf cake days and straw so ate 3 gram of the albino and i couldn't walk and the visuals were amazing life changing


Edited by cyberthreat (02/13/08 11:04 AM)


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