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Offlinethedoc8
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Clone a Clone
    #7443129 - 09/23/07 07:52 AM (16 years, 4 months ago)

I have a master lc that was a clone of a big fruit.

I now have a grow of that lc which made very large fruits.

Is it good/bad to clone a clone...? or just stick with the master lc.

Thanks


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OfflineAndrew47
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Re: Clone a Clone [Re: thedoc8]
    #7443747 - 09/23/07 11:02 AM (16 years, 4 months ago)

It will be fine for now. It's the same as a grain to grain transfer. Eventually you experience some genetic breakdown but that takes a lot more cloning than you are doing.


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Offlinethedoc8
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Re: Clone a Clone [Re: Andrew47]
    #7443852 - 09/23/07 11:42 AM (16 years, 4 months ago)

Thanks


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Invisibleem_bre_O
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Re: Clone a Clone [Re: Andrew47]
    #7443873 - 09/23/07 11:53 AM (16 years, 4 months ago)

Quote:

Andrew47 said:
It will be fine for now. It's the same as a grain to grain transfer. Eventually you experience some genetic breakdown but that takes a lot more cloning than you are doing.




senescence-Mushroom Cultivator page35-
......Senescent mycelium, like any aged plant or animal, is far less vigorous and fertile than it's counterpart.


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OfflineAndrew47
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Re: Clone a Clone [Re: em_bre_O]
    #7444600 - 09/23/07 04:02 PM (16 years, 4 months ago)

exactly.

He's very far from senescence


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Invisibleem_bre_O
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Re: Clone a Clone [Re: Andrew47]
    #7444700 - 09/23/07 04:33 PM (16 years, 4 months ago)

yeah,i know. Jus' puttin' knowledge.


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Offlinemilkman
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Re: Clone a Clone [Re: em_bre_O]
    #7540502 - 10/20/07 07:22 PM (16 years, 3 months ago)

what if say you grow for years and just keep cloning the sexiest looking fruit in each grow will your mushy quality go up or down


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Offline2FiNiTe
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Re: Clone a Clone [Re: milkman]
    #7540572 - 10/20/07 07:44 PM (16 years, 3 months ago)

Gene Degradation.


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Invisibleorchidfanatic
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Re: Clone a Clone [Re: 2FiNiTe]
    #7540733 - 10/20/07 08:33 PM (16 years, 3 months ago)

what if you use the spores from said shroom isolate for best rhizomorphs and then grow from them .. they should be mostly on the large side like the parent the spores came from?


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OfflineRogerRabbitM
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Re: Clone a Clone [Re: orchidfanatic]
    #7541746 - 10/21/07 07:33 AM (16 years, 3 months ago)

Quote:

It will be fine for now. It's the same as a grain to grain transfer.




Not quite. A grain to grain transfer is done with young, rapidly growing mycelium. In no way is that the same as cloning. I would not clone a clone. It isn't necessary. If you have young cell lines, simply store them in the refrigerator for years and keep using them.
RR


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OfflineMushroomGeeks
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Re: Clone a Clone [Re: RogerRabbit]
    #7560282 - 10/25/07 06:44 PM (16 years, 3 months ago)

That is fine to clone a clone. Strain degeneration happens when you clone around 32-100 times...I forget the exact number but it is a lot.


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OfflineRogerRabbitM
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Re: Clone a Clone [Re: MushroomGeeks]
    #7561017 - 10/25/07 10:05 PM (16 years, 3 months ago)

Wrong. Try 3 and performance is substantially weaker. That's why we keep fresh mycelium.
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."
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OfflineSkeeblix
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Re: Clone a Clone [Re: RogerRabbit]
    #7561406 - 10/25/07 11:54 PM (16 years, 3 months ago)

So how can you keep an isolate going on continually if you use most of it, then clone it back out to another dish? From what I hear, you can create a fresh clone off a fruitbody from grown out isolates, but isn't the end result of this the exact same thing as cloning out the isolate in the first place?

This has been something puzzling me for awhile, so I figure I may as well get it sorted in my head before I ever need to do.


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Premedman1 said:
:lol: I just shat my pants.


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OfflineRogerRabbitM
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Re: Clone a Clone [Re: Skeeblix]
    #7562109 - 10/26/07 08:15 AM (16 years, 3 months ago)

You don't.

You keep the original master culture slant in the refrigerator. When you need mycelium, you take a tiny piece from the culture slant and transfer it to a petri dish, then return the slant to the refrigerator, where the mycelium slowly grows to replace what you cut out. Use the petri dish(es) to start the next batch. By doing this, your master slant slows way down on cell division, thus preserving the culture without senescence.
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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OfflineSkeeblix
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Re: Clone a Clone [Re: RogerRabbit]
    #7562319 - 10/26/07 09:39 AM (16 years, 3 months ago)

Hm. Okay, it seems I was a little backwards on the procedure. I kept sitting here wondering how you guys working with agar avoid senescence if you were growing off the culture constantly.

So how is using a small piece off of a master slant to produce a new dish any different? It's still technically the same organism, just grown out in another location. Does this just serve the purpose of allowing you to work off the new dish, avoiding using up a master culture to preserve its favorable traits without risking senescence through another generation of cloning?

This can be so confusing sometimes.


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Premedman1 said:
:lol: I just shat my pants.


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OfflineRogerRabbitM
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Re: Clone a Clone [Re: Skeeblix]
    #7562354 - 10/26/07 09:50 AM (16 years, 3 months ago)

When the culture slant goes into the refrigerator, it essentially stops cell division, putting it into stasis. This way, you can take it out of the fridge, get a small piece, and then return the slant to the refrigerator. Using this system, a 20 year old culture might only have six weeks or less of growth, thus senenscence isn't a problem.
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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OfflineSkeeblix
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Re: Clone a Clone [Re: RogerRabbit]
    #7562370 - 10/26/07 09:55 AM (16 years, 3 months ago)

yeah, I started getting it after a few minutes of mulling it over in my head. the whole idea is to keep the cell line young, so you use the main culture to grow off into a second, then work off the second.

So you can use the master to produce a second generation, which is still fully viable. You can keep using the master to produce second generation cultures that are for the most part unaffected genetically. Then you just work off the 2nd gen cultures and produce new ones when you need them.

Thanks for helping me understand it, it'll come in handy when I start putting it to use.


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Premedman1 said:
:lol: I just shat my pants.


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InvisibleCheezit
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Registered: 10/19/07
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Re: Clone a Clone [Re: Skeeblix]
    #7562770 - 10/26/07 11:28 AM (16 years, 3 months ago)

streak spores on plate. Isolate each colony to new plate. Grow out, section off the growth that appeals to you to more new plates. Get clean pure cultures. Grow out plates, make a master slant. Label, then test the dishes.

You are only sectioning off growth maybe a total on an inch or 2 at this stage. ..------- 2spores(..) growth----- by the time you get the master culture.

now at the time you have the master made, and a few full pure plates grown out. You can then take that plate, do grain with it, make an LC, dice up and make a bunch more plates with it. What ever. But you always got the back up daddy in the fridge.

After testing it out, you will know whether to throw it out or keep it.


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OfflineSkeeblix
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Re: Clone a Clone [Re: Cheezit]
    #7562809 - 10/26/07 11:33 AM (16 years, 3 months ago)

The actual process of doing the agar is no real mystery, it was more the concept of retaining genetic viability through avoiding senescence that was eluding me.

It took me a while to actually get it, but I'm pretty sure I understand it as much as I need to for now.


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Premedman1 said:
:lol: I just shat my pants.


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InvisibleCheezit
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Re: Clone a Clone [Re: Skeeblix]
    #7562960 - 10/26/07 11:54 AM (16 years, 3 months ago)

I got ya.  Sometimes i just go off to reaffirm it myself you know.  Just posting information for others that might not get it totally.  Forgive me if i insulted you with my simple information :wink:


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