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Offlinefinite
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Registered: 08/28/00
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will a strain repeatedly cloned lose its vigor?
    #329205 - 05/29/01 10:52 PM (16 years, 1 month ago)

A question about the process of cloning an isolated strain with mycelia...
I have found a variant amoung a group of 14 jars which were all inocculated with the same
Gulf Coast print from Homestead. The mycelia in this one jar produces mushrooms totally
unlike what the others produced. They are fat, tall, potent, and twisted looking. I managed
to inocculate more jars with mycelia from this strain (it wasn't really a "sterile" technique, but it seemed to work) and plan to case some of them but use some of them as "clone jars" so I can continue
to keep this strain growing. I know taking a spore print will not mean the resulting mycelia
will in fact be identical to the "parent" mushroom. Cloning with mycelia seems to work, I was
just wondering if the strain will lose its vigor if it is used continuously. I know you can clone
with tissue, but why would that be any different than with a pure strain of mycelia?



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Invisibledimitri211
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Registered: 04/27/01
Posts: 2,248
Re: will a strain repeatedly cloned lose its vigor? [Re: finite]
    #329252 - 05/29/01 11:54 PM (16 years, 1 month ago)

Yes it can lose it's vigor P. Stamets uses the p. value system mean the he only transfer mycelium 10 time from agar to agar--If you use a different substrate each time you will do o.k. finch seed then rye grain then wheat grain etc...

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Invisiblellamabox
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Re: will a strain repeatedly cloned lose its vigor? [Re: dimitri211]
    #5003252 - 12/01/05 06:52 PM (11 years, 6 months ago)

Would there be any way to breed a specific clone back to its parent mycelium to stabilize the strain?


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OfflineRogerRabbitM
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Re: will a strain repeatedly cloned lose its vigor? [Re: llamabox]
    #5006096 - 12/02/05 11:45 AM (11 years, 6 months ago)

Just keep those 'clone jars' in the refrigerator. Only take them out when you need to do a grain to grain transfer. They should last for a year or two that way, then simply do a g2g and keep one of those jars back as a strain master.
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Invisibleagar
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Re: will a strain repeatedly cloned lose its vigor? [Re: llamabox]
    #5012981 - 12/04/05 07:28 AM (11 years, 6 months ago)



You can store G-1 LC in 5, 50 or 100 vials of sterile water (years & years).

Any time you want G-1 pull a tube out of the Fridge & go for it.

Either start a new LC with it, or go to grains, then G2G.


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Offlinemattymonkey
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Re: will a strain repeatedly cloned lose its vigor? [Re: agar]
    #5013063 - 12/04/05 09:45 AM (11 years, 6 months ago)

agar- just put a few cc of LC into a vial of sterile water?


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Invisibleagar
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Re: will a strain repeatedly cloned lose its vigor? [Re: mattymonkey]
    #5015776 - 12/05/05 12:04 AM (11 years, 6 months ago)

Quote:

mattymonkey said:
agar- just put a few cc of LC into a vial of sterile water?




You put the LC solution in a rubber stoppered tube.

Allow the myc to settle to the bottom.


You then aspirate out the solution, leaving the myc in the bottom.

(that removes most nutrients).

You then inject sterile water into the tube.


Then refrigerate that tube.

It will remain viable for years.


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Invisiblemicololo2
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Registered: 11/05/05
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Re: will a strain repeatedly cloned lose its vigor? [Re: agar]
    #5016563 - 12/05/05 05:20 AM (11 years, 6 months ago)

Agar- How do you aspirate out the solution without risk of infections?


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Invisibleagar
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Re: will a strain repeatedly cloned lose its vigor? [Re: micololo2]
    #5016659 - 12/05/05 08:16 AM (11 years, 6 months ago)

Soak a few cottom balls in alcohol. Swab down the surface of what you asperate out of, or inject into. Keep the needle wrapped in that alc soaked cotton ball, so it is not exposed to open air.


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OfflineBlue Helix
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Re: will a strain repeatedly cloned lose its vigor? [Re: agar]
    #5021444 - 12/06/05 02:55 AM (11 years, 6 months ago)

I would recommend asperating through the rubber cap. Just inject the air and suck in succession until the free liquid is out. Removing the rubber cap is asking for trouble unless you have a glove box or flow hood.


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Invisibleagar
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Re: will a strain repeatedly cloned lose its vigor? [Re: Blue Helix]
    #5021713 - 12/06/05 04:08 AM (11 years, 6 months ago)

>>>Removing the rubber cap is asking for trouble unless you have a glove box or flow hood.<<<

True that.


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Offlinerungi
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Registered: 07/11/01
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Re: will a strain repeatedly cloned lose its vigor? [Re: agar]
    #6246408 - 11/04/06 11:19 AM (10 years, 7 months ago)

Buffer the medium at various pH values. This causes many mycelia morphologies to differentiate. A clone from a single pure plate to plates of differing pH will all exhibit different growth morphologies.


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OfflineRogerRabbitM
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Re: will a strain repeatedly cloned lose its vigor? [Re: finite]
    #6248655 - 11/05/06 01:05 AM (10 years, 7 months ago)

This thread has been closed.

Reason:
Let's let this old thread on senescence die and go to heaven.
RR


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