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OfflineFugueRider
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How Many Isolates Do You Make? * 1
    #10056817 - 03/27/09 08:46 PM (15 years, 4 days ago)

So they say a grower is supposed to isolate several strains in order to find one that is just right (good fruiter, potent, etc.).

I am wondering how many different isolates you all come up with off of a species. I guess I should have asked this before I actually began isolating; but I went ahead and came up with five different isolates off of one strain. I still need to grow them out though to see what I have

I am just wondering what others do in order to be really successful with strain isolation.


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May All Beings be Peaceful, Happy and Free

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OfflineRogerRabbitM
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Re: How Many Isolates Do You Make? [Re: FugueRider]
    #10056897 - 03/27/09 08:59 PM (15 years, 4 days ago)

The last time I did a series of isolates, I pulled over 30 from a single swipe of spores.  I could have taken twice that many, but intuitively selected what I thought looked like the best 30.  You need to grab quite a few if you want to really hit the jackpot on at least a couple of them.
RR


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OfflineFugueRider
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Re: How Many Isolates Do You Make? [Re: RogerRabbit]
    #10056934 - 03/27/09 09:07 PM (15 years, 4 days ago)

Wow! Thirty eh? Whelp...it looks like I have some more work ahead of me then.

Boy, that is a lot of grows until a guy can develop that intuitive sense you are talking about. It seems like a pretty long learning curve.

Do you have any insight on how to minimize that learning curve any? Or do I just have to put in the hours doing the grows until I start to get a sense of what looks good and what doesn't.

On the plus side, I substituted for a middle school science class earlier this week. I found a great catalog to get tons of petri dishes for dirt cheap...just in the knick of time evidently.


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Offlinepotatonet
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Re: How Many Isolates Do You Make? [Re: FugueRider]
    #10057064 - 03/27/09 09:32 PM (15 years, 4 days ago)

sustituted for a middle school science class... growing mushrooms... oh man


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In the Quechua language of Peru there are over 1000 words for "potato"

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OfflineRogerRabbitM
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Re: How Many Isolates Do You Make? [Re: potatonet]
    #10057497 - 03/27/09 11:16 PM (15 years, 4 days ago)

I've done presentations at both of the local schools in our small county, showing the middle and high school kids how to grow oyster mushrooms on brf cakes, and how to use the cakes to spawn to straw and newspapers they collect.  Next month, I'm doing an exhibit and workshop at the conservation fair where I'm demonstrating mushroom growing techniques as they relate to using waste material to grow mushrooms on.  I think it's great that mushrooms are finally beginning to enter the national consciousness.
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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Offlinepotatonet
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Re: How Many Isolates Do You Make? [Re: RogerRabbit]
    #10057534 - 03/27/09 11:28 PM (15 years, 4 days ago)

I just think they are cool... something from almost nothing =-)


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In the Quechua language of Peru there are over 1000 words for "potato"

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InvisibleJaComet
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Re: How Many Isolates Do You Make? [Re: potatonet]
    #10058599 - 03/28/09 06:09 AM (15 years, 3 days ago)

“ * * *  earlier this week. I found a great catalog to get tons of petri dishes for dirt cheap...just in the knick of time evidently.”

Ahhhh ---- The sweet vibration of synchronicity.

I see many healthy members of your family.


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InvisibleBlimeyGrimey
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Re: How Many Isolates Do You Make? [Re: potatonet]
    #10062358 - 03/28/09 08:47 PM (15 years, 3 days ago)

Quote:

potatonet said:
I just think they are cool... something from almost nothing =-)




Mushroom mycelium is a little reverse entropy engine. Creating order out of chaos. Then again, that's all life. Biology in all its forms is amazing.

Back on topic though. I prefer to clone rather than do multiple isolations. Multiple isolations is good for mating studies, crossing strains and a lot of other uses but for the beginning cultivator it may be easier to start with multispore inoculation. Then clone the best looking fruit.

Clone wild specimens is a good way to get fresh genetics.


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Edited by BlimeyGrimey (03/28/09 09:00 PM)

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Re: How Many Isolates Do You Make? [Re: BlimeyGrimey]
    #10065581 - 03/29/09 11:38 AM (15 years, 2 days ago)

I have to agree with Blimey. Cloning is a much more surefire method of finding the traits you want as opposed to hoping to get those traits in an isolate. I only use agar now primarily for wild prints or prints picked up in trades to check for contaminants and only isolate down until I see a few sectors. That way I can grow with a little genetic variance and clone the specimen with the traits I want from that grow.

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OfflineRogerRabbitM
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Re: How Many Isolates Do You Make? [Re: FooMan]
    #10066650 - 03/29/09 01:44 PM (15 years, 2 days ago)

Quote:

Cloning is a much more surefire method of finding the traits you want as opposed to hoping to get those traits in an isolate.




I strongly disagree. 

Most strains of a single species are compatible.  That means over time, the various sectors(strains) will merge into a common whole via the process of anastomosis. By cloning, you get a 'composite' of the original multispore inoculation.  By isolating dozens of strains, you'll get a few good producers, a few poor producers, a zero producer or two, and a stellar kick-ass producer or two.  The latter are what you save for future use and toss the rest out.  It's more work, but way more productive.
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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Invisiblefastfred
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Re: How Many Isolates Do You Make? [Re: RogerRabbit]
    #10079431 - 03/31/09 10:18 AM (15 years, 17 hours ago)

Anastomosis is pretty rare and is a usually a dead end for the myc from what I've read.

It would be nice to do some further research on the process.

Personally I like the idea of screening thousands or millions of spores at once using MSI rather than 30 or so using direct isolation prior to growing.


-FF

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OfflineRogerRabbitM
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Re: How Many Isolates Do You Make? [Re: fastfred]
    #10079535 - 03/31/09 10:42 AM (15 years, 16 hours ago)

Quote:

Anastomosis is pretty rare




No way.  It happens on every multispore grow.  Tens of thousands of strains combine into one, or just a few, by the time of full colonization.
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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Invisiblefreespeech
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Re: How Many Isolates Do You Make? [Re: RogerRabbit]
    #10079597 - 03/31/09 10:54 AM (15 years, 16 hours ago)

:nothingtoadd:

If anyone knows a way to follow a thread other than by posting in it, let me know :grin:

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InvisibleJaComet
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Re: How Many Isolates Do You Make? [Re: freespeech]
    #10079688 - 03/31/09 11:12 AM (15 years, 16 hours ago)

Bottom of page : Extra information box : hit Toggle Favorites and find in  your Account at top of every page.

(Darn Quote function isn't for me today)

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Offlinesolumvita
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Re: How Many Isolates Do You Make? [Re: fastfred]
    #10080014 - 03/31/09 12:20 PM (15 years, 15 hours ago)

Quote:

fastfred said:
Anastomosis is pretty rare and is a usually a dead end for the myc from what I've read.

-FF



I agree with RR it happens.  As for only cloning, that depends on what you want to do with the mushrooms.  In a commercial sense you need to select for high yielding strains and need to go through the selection process.


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Invisiblefastfred
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Re: How Many Isolates Do You Make? [Re: solumvita]
    #10081188 - 03/31/09 03:31 PM (15 years, 12 hours ago)

Just looked up the easiest to find paper and it listed 1.6-1.3 per cm.  However this doesn't necessarily mean that nuclei are exchanged or that they percolate any distance.

Here is a flowchart of what generally happens when myc meets...



Even if some fruits are a combination of genetics it's easy to separate them on plates and you've still screened hundreds of thousands in one round vs. a few dozen.

RR deals with practical aspects, so listen to his advice.  I only do research and don't give a damn about many aspects.  To me screening large numbers is essential.


-FF


-FF

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InvisibleBlimeyGrimey
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Re: How Many Isolates Do You Make? [Re: fastfred]
    #10084298 - 03/31/09 10:51 PM (15 years, 4 hours ago)

Quote:

fastfred said:
Even if some fruits are a combination of genetics it's easy to separate them on plates and you've still screened hundreds of thousands in one round vs. a few dozen.




That was my reasoning behind cloning. I've had very few clones sector on agar.

I have noticed that my clones of wild friscosas seem to sector more than any other species I've worked with.

Does that mean that the mycelium reverses anastomosis and splits into separate entities again?

I should know most of this since I'm currently taking a Molecular Biology Methods course but we seem to be focusing on bacteria and haven't went into fungi at all.


--------------------
Message me for free microscopy services on Psilocybe, Panaeolus, and Gymnopilus species.

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OfflineRogerRabbitM
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Re: How Many Isolates Do You Make? [Re: BlimeyGrimey]
    #10084978 - 04/01/09 01:38 AM (15 years, 2 hours ago)

Quote:


Does that mean that the mycelium reverses anastomosis and splits into separate entities again?




DNA testing from various parts of wild fruiting bodies has shown that more than one pairing of hyphae can be present in the same mushroom.  In other words, a single mushroom can have multiple strains present.  I've seen it with multispore inoculated grows too, which is basically what wild mushrooms are.
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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Offlinepotatonet
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Re: How Many Isolates Do You Make? [Re: RogerRabbit]
    #10085075 - 04/01/09 02:07 AM (15 years, 1 hour ago)

this would explain why we have 1000 strains of cubensis, because a single mushroom represents multiple strains.

kinda like getting an albino something or a PE off of a B+ grow or something.

makes sense as far as isolation goes.


--------------------
In the Quechua language of Peru there are over 1000 words for "potato"

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InvisibleBlimeyGrimey
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Re: How Many Isolates Do You Make? [Re: potatonet]
    #10085202 - 04/01/09 03:21 AM (15 years, 17 minutes ago)

Anastomosis seems to be a good mechanism to help ensure future generations have genetic diversity.

When a wild mushroom with multiple strains produces spores it will most likely drop spores with a mix of the genetics of both strains.

While a mushroom grown from a single mating of 2 hyphae will only disperse its own (possibly inferior) genes. This is good in a breeding sense for cultivators but bad for genetic diversity.


--------------------
Message me for free microscopy services on Psilocybe, Panaeolus, and Gymnopilus species.

Looking for wild Panaeolus cinctulus and Panaeolus olivaceus prints.

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