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Invisiblebosch
Registered: 01/16/09
Posts: 92
Decisions about isolates
    #10215449 - 04/22/09 06:54 PM (14 years, 11 months ago)

Assuming one uses MS on their first run of a species, planning to isolate strains and run test grows on each isolate from the most promising fruit, what are your personal preferences on choosing which fruit to clone from?

Obviously speed, size, cluster formation, etc are very important, but what I am mainly wondering about is in respect to number of flushes. Seems like the best way to get a strain that produces many would be to choose a fruit from one of the later flushes, correct? But then of course you do not know ahead of time how many you'll get, or if the fruits will be at all good that far along compared to those in the first flushes.

Thoughts?

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OfflineRogerRabbitM
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Re: Decisions about isolates [Re: bosch]
    #10216416 - 04/22/09 09:10 PM (14 years, 11 months ago)

If you're isolating strains, you don't clone at any time.  You mark all your strains and put them in test tubes under refrigeration.  Later, when you determine the qualities you desire from what you stated above, you pull that particular and WELL-MARKED strain out of storage and use it.  Make fresh slants every year in order to keep juvenile mycelium under refrigeration at all times.  Cloning captures mycelium that is already a few months old, thus defeating the purpose of strain isolation.
RR


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Offlinepotatonet
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Re: Decisions about isolates [Re: RogerRabbit]
    #10217009 - 04/22/09 10:38 PM (14 years, 11 months ago)

so you have to make a MS without knowing the fruiting properties?

I guess when you do the BRF cakes you could take a piece of the same section already on a plate and put it onto new agar while the jar grew?

same age myc though... seems like you are putting a lot of work into something that might not work


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Edited by potatonet (04/22/09 10:39 PM)

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Invisiblebosch
Registered: 01/16/09
Posts: 92
Re: Decisions about isolates [Re: potatonet]
    #10217123 - 04/22/09 10:57 PM (14 years, 11 months ago)

Apoligies; I think I used kind of incorrect/misleading terminology, perhaps.

My normal plan of attack is to go multispore on some tray projects, from there use tissue from the best fruits and isolate strains from its interior tissue, perform test grows on those strains to dertermine which I like best, and from there keep that perferred strain for long-term storage, to be used in more wider-scale cultivation techniques, usually straw logs or real wooden logs via plug spawn.

So, it's pretty much openly suggested to keep all isolated strains in potentially long-term storage until one or a few ideals are chosen from that group.

Still, and I could be mistaken on this, but I tend to like to begin my strain isolation projects from successful fruits rather than going from spores directly to agar and arbitrarily selecting strains based on luck and appearance from there before the test grow on the individual isolation stage. Not certain if there is any merit in this or not, but it seems reasonable that strains present to produce desired fruits are more likely to produce more alike in that vein than nearly randomly-chosen strains.

That could be foolish, and if so, it'd be good to know, but otherwise I'd like to hear people's general opinions on how they rate the performance of their isolates, and/or choose fruits from which to attempt to isolate strains from.

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OfflineRogerRabbitM
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Re: Decisions about isolates [Re: bosch]
    #10239149 - 04/26/09 09:02 PM (14 years, 10 months ago)

Quote:

but I tend to like to begin my strain isolation projects from successful fruits rather than going from spores




Usually, but not always, mushrooms grown indoors are isolated strains.  I've noticed exceptions to this rule, but it's true over 95% of the time, so you're not going to get many isolates from cloning.

If you want to isolate, do it from spores and grow each isolate to the fruiting stage.  That's how it's done.  Yes, it's more work.
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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OfflineVoodoo486
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Re: Decisions about isolates [Re: RogerRabbit]
    #10240964 - 04/27/09 03:03 AM (14 years, 10 months ago)

It seems to me that the only way to get the most consistent characteristics from fruits is either strain isolation on agar, or via cloning.

Bosch, your method of picking a few fruits and cloning them is- from what I have read- exactly how you're supposed to do it. On the other hand, RogerRabbit is correct in the statement that your cloned mycelium will start at the age of the cloned fruit.

Both get the same result, but agar is a better method.


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Offlinesheikofshiitake
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Re: Decisions about isolates *DELETED* [Re: Voodoo486]
    #10241319 - 04/27/09 06:54 AM (14 years, 10 months ago)

Post deleted by sheikofshiitake

Reason for deletion: .


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Invisiblebosch
Registered: 01/16/09
Posts: 92
Re: Decisions about isolates [Re: sheikofshiitake]
    #10249599 - 04/28/09 01:51 PM (14 years, 10 months ago)

Yeah, good point RR, and the rest (:

About a day after I made that post and re-referenced some stuff in the Stamets books I felt pretty dumb when I was reminded of the p-system, hah. While you may be more likely to get desirable traits trying to isolate from a fruit biopsy, yeah, it's way further along the road to senescence, and both methods require probably about the same number of experimental grows anyway. Kind of embarrassing, hah.

Thanks for the input however!

A somewhat related question however which I am curious about: So, say I am isolating from spores on agar plates. I take small samples, transfer to new plates etc etc to reduce the number of sectors/strains progressively, do the experimental grows once isolates are gotten, and determine which I like best, but let's say I keep all the old dishes I transferred from down the line in cold storage. Question is this: Say I am looking at the very last dish which still had sectoring from which I got my desired isolate... Are the other sectors on this plate as well more likely to have the desired traits?

Never would I do this, and I'd never recommend anyone doing so, as it'd be pointless and stupid, but I curious on the genetics level. Each strain is a combination two 'genders', if memory serves? My forgetfulness regarding terminology makes explaining my thoughts more difficult... But, the other sectors would probably be more likely to have at least one of these, I would imagine, and thus be more likely to have similar traits?

Seems to be reasons for me to think it reasonable for there to be some increased likelihood AND no increased likelihood, hah. Maybe someone can clear it up a little for me.

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OfflineRogerRabbitM
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Re: Decisions about isolates [Re: bosch]
    #10249760 - 04/28/09 02:18 PM (14 years, 10 months ago)

Each strain is a pairing of two compatible hyphae.  Other sectors on the same plate are made up of pairings of different hyphae.  Characteristics would not be identical.
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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