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OfflineFGL
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differences betwen strain isolation by transfers and by cloning?
    #5831095 - 07/07/06 11:13 AM (17 years, 8 months ago)

If I take a clone from a multispore growing am I getting the same strain qualities than doing isolations on petri dishes? :rolleyes:

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OfflineRogerRabbitM
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Re: differences betwen strain isolation by transfers and by cloning? [Re: FGL]
    #5831103 - 07/07/06 11:15 AM (17 years, 8 months ago)

No.


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OfflineFGL
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Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #5831131 - 07/07/06 11:24 AM (17 years, 8 months ago)

Quote:

RogerRabbit said:
No.




So for getting the best of a strain is it better to do transfers?

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OfflineRogerRabbitM
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Re: differences betwen strain isolation by transfers and by cloning? [Re: FGL] * 1
    #5831155 - 07/07/06 11:30 AM (17 years, 8 months ago)

One swipe of spores on agar will yield hundreds of strains. By selecting a dozen or so of the best rhizomorphic strains and fruiting each one separately, you can find the super performer that will cover every spot of your casing layer with healthy pins. It might take hundreds of multispore grows to find that strain, if ever, because multispore inoculated substrates usually end up with only one or two strains by the time they fruit because they've all combined. (anastomosis)

This means the good fruiting and potent strains combine with the poor fruiting and bunk strains. You never know what you're going to get. Isolate on agar, then fruit separately and test each strain. When you find the best one, you can keep master slants in the refrigerator and grow that isolate forever.
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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OfflineFGL
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Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #5831356 - 07/07/06 12:20 PM (17 years, 8 months ago)

Got it! :thumbup:
Many thanks for your help

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InvisiblePsychoslut
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Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #5831383 - 07/07/06 12:25 PM (17 years, 8 months ago)

How do you avoid not running out of original master slants? How do you keep growing out the same isolate and avoid senescence and all that?


Ive read that you cant do too many transfers before the isolate goes bunk. So how do you not run out of "P1"??


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[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]

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Invisiblefastfred
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Re: differences betwen strain isolation by transfers and by cloning? [Re: Psychoslut]
    #5832740 - 07/07/06 05:53 PM (17 years, 8 months ago)

> One swipe of spores on agar will yield hundreds of strains.

Come on RR, stop calling each individual a "strain". They're sibling individuals and share 50% of their DNA, that hardly qualifies calling each of them a strain.

> How do you avoid not running out of original master slants?

You only take a speck from your master slant and then put it back in the fridge. You only take a tiny bit each time and you grow it out to however much you need. You also keep active cultures going if you need to frequently access that sample.


-FF

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OfflineRogerRabbitM
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Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred] * 1
    #5833313 - 07/07/06 08:50 PM (17 years, 8 months ago)

Two compatible hyphae pairing up and producing a dykaryon is the mycological definition of a strain. The OMC use of the word strain to denote the geographical area the parent stock came from is marketing driven.
RR

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Offlinehyphae
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Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred]
    #5833318 - 07/07/06 08:52 PM (17 years, 8 months ago)

Quote:

fastfred said:
> One swipe of spores on agar will yield hundreds of strains.

Come on RR, stop calling each individual a "strain". They're sibling individuals and share 50% of their DNA, that hardly qualifies calling each of them a strain.

> How do you avoid not running out of original master slants?

You only take a speck from your master slant and then put it back in the fridge. You only take a tiny bit each time and you grow it out to however much you need. You also keep active cultures going if you need to frequently access that sample.


-FF



True fastfreddy they are indeed all substrains, I believe thats what RR means but I'm glad you caught that shows good solid mycological understanding.


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Invisiblefastfred
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Re: differences betwen strain isolation by transfers and by cloning? [Re: hyphae]
    #5833581 - 07/07/06 10:21 PM (17 years, 8 months ago)

Strain: Biology A group of organisms of the same species, having distinctive characteristics but not usually considered a separate breed or variety.

Strain: (biology) a group of organisms within a species that differ in trivial ways from similar groups.

Strain: A strain is sometimes used to indicate a group of plants with similar (but not identical) appearance and/or properties. The term has no official status.


> Two compatible hyphae pairing up and producing a dykaryon is the mycological definition of a strain.

References?

The mating of two monokaryons from the same parent, as is referred to in this post, represents the absolute minimum of genetic variation resulting from the completion of the sexual cycle. It isn't a "strain" and even calling it a "substrain" is being VERY generous.


-FF

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OfflineRogerRabbitM
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Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred] * 1
    #5834378 - 07/08/06 05:43 AM (17 years, 8 months ago)

I've given the mycological definition of a strain above. I won't argue semantics with you because your old biology book says something different. What I gave is the accepted mycological definition of a strain. Perhaps you can write letters to Stamets, Holliday, Volk, etc., and tell them they're all wrong.
RR


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Offlinehyphae
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Re: differences betwen strain isolation by transfers and by cloning? [Re: fastfred]
    #5834432 - 07/08/06 07:34 AM (17 years, 8 months ago)

Quote:

fastfred said:
Strain: Biology A group of organisms of the same species, having distinctive characteristics but not usually considered a separate breed or variety.

Strain: (biology) a group of organisms within a species that differ in trivial ways from similar groups.

Strain: A strain is sometimes used to indicate a group of plants with similar (but not identical) appearance and/or properties. The term has no official status.


> Two compatible hyphae pairing up and producing a dykaryon is the mycological definition of a strain.

References?

The mating of two monokaryons from the same parent, as is referred to in this post, represents the absolute minimum of genetic variation resulting from the completion of the sexual cycle. It isn't a "strain" and even calling it a "substrain" is being VERY generous.


-FF




fastfreddy we work with different strains ie; PESA, PESH, Amazonians, B+, etc; they all have different charateristics that remain consistant. Now when we take one of thoses strains and do a multispore we create substrains I don't care how you interpret what you read this is simple and easy for anyone/everyone to understand. Were not trying to get too technical here, substrain is a well understood term used in mycology that at least gets us all on the same page. TC bro


--------------------
Getting the most out of your casings!, A pinning strategy.
Oyster Shell "Flour" $2 for 1lb. a hell of a deal :wink:
Not what is overlay but rather what overlay is
Gas Exchange vs. FAE

"We all have priorities. I used a closet once setup a nice little lab trouble was all the shit that was in there ended up in the bedroom that pissed off the GF then I ended up dumping her as she was getting in the way of my sterile culture technique! Ya I got priorities too!!!"

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Offlinezeegos
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Re: differences betwen strain isolation by transfers and by cloning? [Re: hyphae]
    #6293261 - 11/17/06 08:50 AM (17 years, 4 months ago)

I think theres something i dont quite understand. RR, iv read your write-up on strain isolation on mycotopia and understand that you select different strains from multispore on agar to find the best performing strain. first off your looking for a fruiting strain (of course  :rolleyes:), one which produces large fruits and good pinsets. but i dont understand how this cant be achieved at the same standard as cloning? i mean, isnt cloning different healthy shrooms the same as isolating different strains on an agar dish?

woops, i forgot that only 1 or 2 dominant strains make up the fruits, nevermind  :tongue:

Edited by zeegos (11/17/06 08:53 AM)

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OfflineBluemoondreamer
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Re: differences betwen strain isolation by transfers and by cloning? [Re: zeegos]
    #6293375 - 11/17/06 09:28 AM (17 years, 4 months ago)

And strains also share more than 50% of their genes too.

"We found the average nucleotide identity (ANI) of the shared genes between two strains to be a robust means to compare genetic relatedness among strains, and that ANI values of ≈94% corresponded to the traditional 70% DNA–DNA reassociation standard of the current species definition. At the 94% ANI cutoff, current species includes only moderately homogeneous strains, e.g., most of the >4-Mb genomes share only 65–90% of their genes, apparently as a result of the strains having evolved in different ecological settings."

Edited by Bluemoondreamer (11/17/06 09:54 AM)

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OfflineBluemoondreamer
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Re: differences betwen strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #6293400 - 11/17/06 09:41 AM (17 years, 4 months ago)

To be clearer - when multispore wipes from the same parents are used, they typically retain high 90's % of the same genes. Very little if at all genetic drift.

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OfflineBluemoondreamer
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Re: differences betwen strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #6293417 - 11/17/06 09:49 AM (17 years, 4 months ago)

strain -

1. The collective descendants of a common ancestor; a race, stock, line, or breed.
2. Any of the various lines of ancestry united in an individual or a family; ancestry or lineage.
3. Biology A group of organisms of the same species, having distinctive characteristics but not usually considered a separate breed or variety: a superior strain of wheat; a smooth strain of bacteria.

There is RR's reference if u really need one.. I think I learned it back in HS.

*edit a perfect example would be compatable hyphae from PF albino and PE pairing - another example would be compatible hyphae from spores within the same PE syringe pairing up. Regardless of the level of DNA similarities - inbreeding or outbreading results in new strains. 1% of genetic drift may not look like worlds apart on paper, but it is worlds apart on the genome level.

Edited by Bluemoondreamer (11/17/06 10:13 AM)

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OfflineMycoMaster83
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Re: differences betwen strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #6293624 - 11/17/06 11:18 AM (17 years, 4 months ago)

^Interesting

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Invisiblecreamcorn
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Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293684 - 11/17/06 11:41 AM (17 years, 4 months ago)

Quote:

RogerRabbit said:
One swipe of spores on agar will yield hundreds of strains.  By selecting a dozen or so of the best rhizomorphic strains and fruiting each one separately, you can find the super performer that will cover every spot of your casing layer with healthy pins.  It might take hundreds of multispore grows to find that strain, if ever, because multispore inoculated substrates usually end up with only one or two strains by the time they fruit because they've all combined. (anastomosis)

This means the good fruiting and potent strains combine with the poor fruiting and bunk strains.  You never know what you're going to get.  Isolate on agar, then fruit separately and test each strain.  When you find the best one, you can keep master slants in the refrigerator and grow that isolate forever.
RR




My apologies to responding to part of this thread from months ago, but somebody else bumped it and has me thinking again, I still don't understand this... and I remember talking with RR about it a while back.

I still don't understand what the fundamental difference in the "amount" of varied genes would be between a pure isolate, and a clone.  It still seems to me that both (assuming proper procedure) are isolated strains, one single unified organism, with one single set of genetic material.  So at least it SEEMS to me that both are a different means to the same end.

The difference is that multispore on agar yields many strains (substrains, however you want to word it), so you have many to pursue for isolation, and several to choose from.  By looking for rhizomorphic growth, you have a pretty good idea you'll have a fruiter, but its not a gauruntee.  You have no idea how good of a fruiter it is, until you have fruited it.  You may need to try several, but again you have several to choose from, so one is bound to stand out above the others.  Still, you're not garunteed what you have is near the "best"... you may have been unlucky and got a bunch of crappy strains to choose from - but you don't know until a considerable investment of time, materials, and effort has taken place and you've fruited each and compared.

On the other hand, cloning doesn't leave you with that choice.  On a single grow, that's it.. through anastamosis you've likely narrowed your substrate down to a few (sub)strains, and it still has to do with chance, but your choices are now limited.  So you clone a fruitbody... at least you *know* you're taking genetic material from something capable of fruiting, because the proof is in hand already.  If you happen to get lucky and not only grew a good performer by chance, but chose a good performer, then haven't you reached the game goal? 

Seems to me you're tackling the same problem, only starting at the beginning (agar) versus starting at the end (cloning) and there's pros and cons to each. 

I guess I still don't understand or know for sure that a clone is truly "isolated"... you might have a mix of things that made it up due to the anastamosis... but couldn't that also occur right from inoculation on a petri?  I know you can observe two sectors coming together and a third one emerging, I've seen it.  Isn't the same thing that happens as your particular grow's substrate "joins up" into a single body happening there, just at an earlier stage?  Are we concerned that the third sector came from two others, and didn't purely emerge from spores?  (If so, that's news to me about how you should choose sectors to pursue on agar!) 

At any rate, my 2 cents is I've done it both ways, and cloning really works for me.  (In fact I'm gonna get some plates going next week, I've had a sleeve of petris left over untouched for over a year now and enough agar around to make a thanksgiving jell-o mold with haha, might as well play around :wink:)  But from my personal experiences, while cloning isn't necessarily as reliable to get off the ground with, and the choices are limited to basically the "result" of any given fruiting tray, it has produced the best results for me in the long run, with less effort and specialty materials involved I might add.  The LC cloning I describe in my other thread is basically the conclusion I came to after doing things both ways, its a method that requires little skill, few materials, and even with the "chance" involved in getting a quality specimen to clone, and found that supposed chance at least for me, to *always* be in my favor, with noticably more abundant and potent crops coming from each and any clone I've ever taken, versus starting from multispore.  Maybe I'm alone on this and just lucky with some good mushroom karma? :smile:

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OfflineEraserhead
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Re: differences betwen strain isolation by transfers and by cloning? [Re: MycoMaster83]
    #6293685 - 11/17/06 11:41 AM (17 years, 4 months ago)

Why not call the seperate isolates "phenotypes" ? like with MJ, each strain has many variant phenotypes.


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Invisiblecreamcorn
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Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead]
    #6293696 - 11/17/06 11:46 AM (17 years, 4 months ago)

Quote:

Eraserhead said:
Why not call the seperate isolates "phenotypes" ? like with MJ, each strain has many variant phenotypes.




I believe "phenotype" would be reserved for describing characteristics of an actual fruitbody.  For example, I have a culture that produces mushrooms with dimples in their caps (like the pic in my avatar)... I'd consider the "dimple cap" culture I have, a phenotype.  It all points back to my confusion on my long rant above, is a phenotype then, a single set of genetics, or is it a description of a single set of characteristics?  Is it truly an isolated organism?  Would that explain why we have all these words to describe it? :wink:

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