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OfflineMeneerCactus
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Any-one done this before ? Atlantis Cloning ;)
    #1404810 - 03/24/03 05:03 AM (13 years, 8 months ago)

High,

Today I tried to do some cloning of sclerotia.

Last week I had a morning dream about it and so I was very motivated to work it out. I used  some commercial sold sclerotia to work with.

Here is what I did according to my dream:
I made a nice sterile bath/container to work in filled with a sterilized water and h202 solution.
I immeaditaly dropped my sterile tools in it.

All the cloning happened under water/h202, and the wet mycelium was quickly tranferred to the sterile grow-medium.

Working this way felt very save, was very easy and fun to do/try.

If it is something new, what I doubt, let me hear so!

In a few days I will post the results. Hoping the sclerotia is still fresh enough. . .

:laugh:



   


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InvisibleOlgualion
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: MeneerCactus]
    #1404864 - 03/24/03 05:28 AM (13 years, 8 months ago)

You can even clone dried sclerotia by allowing it to rehydrate in H2O.  Then take a chunk and spear it with your scalpel.  Let this bubble away in a 50% H2O2 /H2O sol'n for a minute, and from here, go to peroxidated agar.

I don't know if it will work with the skipping of the agar step.    You may find that the mycelium won't jump off of the stone and onto the substrate.  It can be stubborn even with agar.  Nevertheless, best of luck to you. :smile: 


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OfflineAnnoA
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: MeneerCactus]
    #1425260 - 04/03/03 03:49 AM (13 years, 8 months ago)

And?


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OfflineMeneerCactus
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: Anno]
    #1425273 - 04/03/03 03:57 AM (13 years, 8 months ago)

High Anno,



This picture was taken a few days ago, in the meantime the mycelium has grown very nice. I did not get any contamination.

I was so happy about it that I also did some Atlantas Cloning on Tampanensis.It feels very save to work this way under a protecting layer.


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OfflineMeneerCactus
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: Olgualion]
    #1425442 - 04/03/03 06:36 AM (13 years, 8 months ago)

High,

I will inoculate some mycelium to grasseeds tommorow and see if it will colonize.
Ofcoarse I keep you updated :smile: 


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OfflinePaid
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: MeneerCactus]
    #1425470 - 04/03/03 07:20 AM (13 years, 8 months ago)

Good luck :-) whats the make up of the cloning water?


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OfflineMeneerCactus
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: Paid]
    #1428130 - 04/04/03 08:08 AM (13 years, 8 months ago)

Quote:

Good luck :-) whats the make up of the cloning water?



I used water/brew-body(malto-dextrines) mixeture.
I added 4% of the brew-body to hot water, mixed it very well, filtered it and sterilezed it for 20 minutes in the PC.


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OfflinePaid
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: MeneerCactus]
    #1428141 - 04/04/03 08:26 AM (13 years, 8 months ago)

Ah no peroxide then?maybe thats were im going wrong as i cant seem to get a good liquid culture system.I'll try your mix next t.x


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OfflineMeneerCactus
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: Paid]
    #1428155 - 04/04/03 08:38 AM (13 years, 8 months ago)

Oeps, I totally forgot to mention the 1 % H202. I added the H202 to the solution after the clone was added and before the jar was closed.

And as mentioned in the top post I used H202 too at the cloning table :smile:

BTW Test-jar is inoculated :smile:


   


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Offlinethe spiral
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: MeneerCactus]
    #1431303 - 04/05/03 02:31 PM (13 years, 8 months ago)

Quote:

I added the H202 to the solution after the clone was added and before the jar was closed.




that's weird, usually i add the h2o2 to the jar after i close it.


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OfflineMeneerCactus
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Re: Any-one done this before ? Atlantis Cloning ;) [Re: the spiral]
    #1432791 - 04/06/03 03:17 AM (13 years, 8 months ago)

Just trying out .....


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