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OfflineTruffleRuffler
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How many transfers is too many?
    #26120180 - 07/20/19 06:25 PM (1 year, 7 months ago)

I only ask this because I've seen conflicting information in different sections of the forum. Some places I've seen it takes close to a dozen transfers to attempt a monoculture with good genetics. But other places on here I've seen it said by 3 or 4 transfers a culture originating from ms syringe begins to senesce and it begins to deteriorate. So what is the real answer? If someone could chime in that would be great.


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OfflineLyh
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Re: How many transfers is too many? [Re: TruffleRuffler]
    #26120208 - 07/20/19 06:39 PM (1 year, 7 months ago)



--------------------
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OfflineSvetaketu
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Re: How many transfers is too many? [Re: Lyh]
    #26120345 - 07/20/19 07:55 PM (1 year, 7 months ago)

Just do as many as you need until it's clean.

A monoculture is not necessary.

Quote:

Lyh said:
https://www.shroomery.org/forums/showflat.php/Number/25203525#25203525




Monokaryotic mycelium is something else entirely.


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Edited by Svetaketu (07/20/19 07:56 PM)


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InvisiblePastywhyteM
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Re: How many transfers is too many? [Re: Svetaketu]
    #26120350 - 07/20/19 08:00 PM (1 year, 7 months ago)

Don’t worry about getting a true fruiting isolate. You will need a dozen to find a good one and have to fruit all of them to see which one is good. Just clone.


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Offlineverum subsequentis
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Re: How many transfers is too many? [Re: Svetaketu]
    #26120355 - 07/20/19 08:05 PM (1 year, 7 months ago)

it would appear you didn't read the whole thing. it starts talking about monokaryons but then talks about how many transfers to achieve a single sector isolate (aka monoculture).


Without serial dilution, the fastest I've ever achieved a mono is around 30 transfers. I've also transferred up to 48 times without finding an iso. It all depends on how many sets of genetics you start with and how small your transfers are.


As to whether a mono is needed.... That's still very much under debate. Of course it isn't completely necessary but it's yet to be (and may never be) proven whether or not a mono culture is possible of knocking the socks off of anything from ms. Most folks never achieve a true mono and even if they do, they don't test it enough or test enough different monos to really find the truth. I actually just busted out a bunch of isolated genetics from cold storage and am in the process of taking back up the experiments. They are penis envy though so it's going to be hard to compare to standard genetics.


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OfflineLyh
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Re: How many transfers is too many? [Re: Svetaketu]
    #26120360 - 07/20/19 08:06 PM (1 year, 7 months ago)

Quote:

Svetaketu said:
Just do as many as you need until it's clean.

A monoculture is not necessary.

Quote:

Lyh said:
https://www.shroomery.org/forums/showflat.php/Number/25203525#25203525




Monokaryotic mycelium is something else entirely.




Ahh you're right I'm sorry. Got stoned and mixed it up haha


--------------------
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InvisiblePastywhyteM
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Re: How many transfers is too many? [Re: Lyh]
    #26120393 - 07/20/19 08:31 PM (1 year, 7 months ago)

Alan Rockefeller believes a clone is for all intents and purposes a single culture. Just because it’s not a true fruiting isolate doesn’t mean the genetic code isn’t stable according to him and other mycologists. Sidnee Obersinglton says the same. If you really want a true fruiting isolate you need to make it with monokaryons. Unless you got a scope there is no point in trying to do that.


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Offlinek5hd2y
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Re: How many transfers is too many? [Re: Pastywhyte]
    #26120395 - 07/20/19 08:34 PM (1 year, 7 months ago)

:nodofunderstanding:


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OfflineSvetaketu
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Re: How many transfers is too many? [Re: Lyh]
    #26120412 - 07/20/19 08:44 PM (1 year, 7 months ago)

Quote:

Lyh said:
Quote:

Svetaketu said:
Just do as many as you need until it's clean.

A monoculture is not necessary.

Quote:

Lyh said:
https://www.shroomery.org/forums/showflat.php/Number/25203525#25203525




Monokaryotic mycelium is something else entirely.




Ahh you're right I'm sorry. Got stoned and mixed it up haha




All good.

As verum pointed out, it does actually have something relevant to the discussion :rofl:

Quote:

bodhisatta said:

It takes doezens of transfers to even get to the point of sectoring and easily 20-40+ to get an isolate. And even more if you put an entire drop of spore solution on a dish





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Offlineverum subsequentis
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Re: How many transfers is too many? [Re: Svetaketu]
    #26120446 - 07/20/19 09:09 PM (1 year, 7 months ago)

i'm not going to disagree with you or Alan or the other guy but... I'm not sure I completely agree. I understand in some ways but would not entirely agree in others. And then it would also entirely depend on the particular clone.

I can only argue in theory as I haven't had enough experience with isolates but I do know that clones can contain varied genetics with varied phenotypes and varied maturation rates. Anastomosis could potentially homogenize all compatible genetics but there could still be non compatible genetics present in a clone.

I don't really think going after monokaryons would be entirely necessary to find the value of an isolate. Simply testing enough sets of genetics without allowing them to combine with genetics around them offers a mycologists a potential opportunity to find the bees knees and slant it.

I do agree that cloning is amazing and by far the fastest way to achieve a kick ass and reliable culture.


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Re: How many transfers is too many? [Re: verum subsequentis]
    #26120502 - 07/20/19 09:46 PM (1 year, 7 months ago)

DNA sequence is a pretty reliable and convincing argument for clones being one set of genetics IMO. That is what they base it on.


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Offlineverum subsequentis
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Re: How many transfers is too many? [Re: Pastywhyte]
    #26120508 - 07/20/19 09:47 PM (1 year, 7 months ago)

interesting indeed. I admittedly don't know jack about dna sequencing.


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OfflineTruffleRuffler
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Re: How many transfers is too many? [Re: verum subsequentis]
    #26120509 - 07/20/19 09:48 PM (1 year, 7 months ago)

Okay, a couple things. 1. Great conversation about the exact tenets and viability of a monoculture. But 2. None of the replies actually answer my question. So if monocultures are unviable. What number of transfers is "good enough" and what number would be too many? 3. 'True Fruiting Isolate' has been mentioned several times but what exactly IS that? Or is it simply and for all intents and purposes a fancy way of saying monoculture?


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Re: How many transfers is too many? [Re: TruffleRuffler]
    #26120519 - 07/20/19 09:54 PM (1 year, 7 months ago)

It’s a monoculture that only ever contained two nuclei


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OfflineSubfinder
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Re: How many transfers is too many? [Re: TruffleRuffler]
    #26120527 - 07/20/19 09:59 PM (1 year, 7 months ago)

Not all isolates will fruit. Some just grow mycillium forever...

You can transfer that till the cows come home, still won't fruit for shit.

That's why multispore to clone is a more efficient way to get a fruiting isolate.


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Offlineverum subsequentis
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Re: How many transfers is too many? [Re: TruffleRuffler]
    #26120538 - 07/20/19 10:05 PM (1 year, 7 months ago)

Quote:

TruffleRuffler said:
Okay, a couple things. 1. Great conversation about the exact tenets and viability of a monoculture. But 2. None of the replies actually answer my question. So if monocultures are unviable. What number of transfers is "good enough" and what number would be too many? 3. 'True Fruiting Isolate' has been mentioned several times but what exactly IS that? Or is it simply and for all intents and purposes a fancy way of saying monoculture?




How many is enough.. transfer until you have a clean culture. It will look nice and organized if it's clean. They won't always look the same but they should look organized.



that picture shows a couple clean cultures that look different from one another.

How much is too much. If you keep transfering you'll end up with a culture that shows no sectors



transfering beyond this point is "too much" simply because you are no longer accomplishing anything but are causing your culture to grow older and weaker by making it grow more.


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OfflineTruffleRuffler
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Re: How many transfers is too many? [Re: verum subsequentis]
    #26120608 - 07/20/19 11:02 PM (1 year, 7 months ago)

Thank you all! Quite the interesting topic imo. Also verum thats some nuts looking myc especially the one at the top. Never seen rhizo curve like that.


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Re: How many transfers is too many? [Re: Pastywhyte]
    #26120688 - 07/21/19 12:01 AM (1 year, 7 months ago)

Quote:

Pastywhyte said:
Alan Rockefeller believes a clone is for all intents and purposes a single culture. Just because it’s not a true fruiting isolate doesn’t mean the genetic code isn’t stable according to him and other mycologists. Sidnee Obersinglton says the same. If you really want a true fruiting isolate you need to make it with monokaryons. Unless you got a scope there is no point in trying to do that.



i still think genetics is in its infancy. with things like HGT being found out and blowing peoples minds when it was. it wouldn't be surprising if they found out in the future new "crazy things" like if DNA has code to change itself into different DNA in some cases. or other such unthinkable things.


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OfflineNef
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Re: How many transfers is too many? [Re: bodhisatta]
    #26120786 - 07/21/19 01:11 AM (1 year, 7 months ago)

this thread and some of the links off it have been an education

thanks fellas :rockon: helped clear up a fair bit for me there


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Re: How many transfers is too many? [Re: Nef]
    #26120803 - 07/21/19 01:18 AM (1 year, 7 months ago)

Quote:

bodhisatta said:
The mushrooms themselves are made of many strains in most cases.

Quote:

bodhisatta said:
Clones are often made of multiple strains their performance may come from the interactions of the contained genetics.

http://www.tandfonline.com/doi/pdf/10.1271/bbb.67.100
Check page 5

Quote:

bodhisatta said:
there's good strains but strains are the resultant of two spores mating. so you can have good or shit strains of any variety(what a vendor calls a strain) a variety is like GT B+ etc... so really it's up to your agar work to obtain a good strain of any of the available varieties


you might have 100s of strains in a single cake if you injected spores into it. some of the mushrooms are made of more than a few strains themselves.

Quote:

bodhisatta said:


here's some nameko fruits that are displaying some white some red and some mixed. the ones in the middle are strains mixing via a process called anastomosis. the resulting fruits are called chimeric




like these nameko mushrooms with some white some red and some mushrooms spotted as the strains both have some phenotype differences manifesting in the ones with multiple strains 










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