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OfflineMycorial
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Transfers vs Cloning
    #10325052 - 05/11/09 11:47 PM (14 years, 9 months ago)

I'm about to begin working with Agar for the first time. Since this is a bit of an advanced technique I just wanted to post a quick question here. :smile: My apologies if this should be in the common area, I leave it to your discretion :smile:

I've seen both of RRs videos regarding both Cloning mushroom tissue and using spore prints with an inoculation loop.

When cloning mushroom tissue, you still have sectoring going on right? Or is the mushroom body the result of a single strain?

Thanks for your time!


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InvisibleShroominit
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Re: Transfers vs Cloning [Re: Mycorial]
    #10325222 - 05/12/09 12:34 AM (14 years, 9 months ago)

You still have sectors MOST of the time I'd say probably more than 99% of the time. Many people will tell you to go from spores because you will have much younger mycelium for your master culture if you do. (Less cell divisions = genetically younger)


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OfflineMycorial
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Re: Transfers vs Cloning [Re: Shroominit]
    #10325973 - 05/12/09 06:59 AM (14 years, 9 months ago)

I've heard RR talk about how things get a bit worn out over time. I'm still using the genetics of the spores I received 2 months ago. I have not taken a fresh sample since then.

What are the disadvantages of continuing at this point with a clone tissue? Will things just grind to a near halt growth wise in the mycellium, less performance/more throwbacks overall?

Thanks Shroominit :smile:


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InvisibleDoc_T
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Re: Transfers vs Cloning [Re: Mycorial]
    #10326112 - 05/12/09 08:04 AM (14 years, 9 months ago)

Two months from spores?
You should be fine.


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OfflineRogerRabbitM
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Re: Transfers vs Cloning [Re: Mycorial]
    #10326248 - 05/12/09 08:57 AM (14 years, 9 months ago)

Quote:

Mycorial said:

When cloning mushroom tissue, you still have sectoring going on right? Or is the mushroom body the result of a single strain?

Thanks for your time!




I just typed this reply out this morning.
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: Transfers vs Cloning [Re: RogerRabbit]
    #10327131 - 05/12/09 12:54 PM (14 years, 9 months ago)

> You still have sectors MOST of the time I'd say probably more than 99% of the time.

IME this is pretty rare.  OTOH myc even from an isolate will sector eventually.  IMHO sectoring isn't a good indication of genetics, and has a lot to do with lifecycle and environmental cues.

All myc of any genetics has the ability to form different sectors with different types of growth.

Some of it may have to do with the technique used.  If you take a cross section or a biopsy you're going to have several different tissue types in your culture.  There's going to be 'skin' cells and probably a couple different types of inner stem tissue cells.  Obviously they're going to have different morphology and are going to sector.

I'm sure that sometimes strands of myc from different sub-strains (or individuals) is going to get caught up in the fruit body production.  And because they experience the same chemical triggers as the rest they will integrate and form a seamless looking fruit body.

In either case cloning technique is going to matter.  If you carefully dissect a stem and take only a small piece of inner stem tissue IME it's pretty rare to get any sectoring unless you grow it out long enough for the tissue to differentiate on it's own.


Quote:

What are the disadvantages of continuing at this point with a clone tissue? Will things just grind to a near halt growth wise in the mycelium, less performance/more throwbacks overall?




It will work just fine for at least a run or two.  I used to take a clone and grow up a quart of LC from it.  I usually took several of the best clones so I had a couple quarts of LC.  I don't think I actually ever used up a full quart and I never bothered re-cloning or something silly like that.


Hopefully most people here are actually interested in a breeding program.  Cloning just to get a good isolate misses most of the point IMHO.  You should clone to get an isolate, see how it performs, then use it's spores to produce a new generation, then repeat.  In that way you can be constantly be improving your strain's genetics.

That's the starting point for an advanced breeding program.  First you want to take your strain and isolate the best genetics you can.  Then by inbreeding or line breeding you work to get the most consistent strain you can.  At that point you will want to out-cross it to another strain with which you've done the same thing.  In that way you isolate the best genetics from several strains then you cross them together in hopes of producing a new strain with the best of the genetics from the two parent strains.


-FF


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OfflineRogerRabbitM
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Re: Transfers vs Cloning [Re: fastfred]
    #10327286 - 05/12/09 01:32 PM (14 years, 9 months ago)

It's also important not to confuse different mycelium types with sectors.  Sectors are distinct genetic individuals, while different mycelium types can be from age, food source, environmental conditions, etc.
RR


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semper in excretia sumus solim profundum variat

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OfflineMycorial
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Re: Transfers vs Cloning [Re: Mycorial]
    #10328832 - 05/12/09 07:04 PM (14 years, 9 months ago)

Very informative, thanks for taking the time to respond :smile:

This is an interesting new chapter for sure!


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