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OfflinePsilotoad
Monke
Registered: 04/23/22
Posts: 9
Last seen: 2 months, 6 days
Re: Dedikaryotization of Cubensis fruit body clones? [Re: Nillion] * 1
    #27748189 - 04/23/22 02:08 PM (1 year, 9 months ago)

Leal-Lara and Egar-Hummel 1982 method worked for me with a Long Ghost isolation and 15 second blend times. I was only able to recover one haplont due to mistakes on my end but the other was there because what wasn't recovered and plated from the solution became dikaryotic again. I am in the process of running it again on a few more strains and will hopefully get symmetrical recovery. Will provide a pictorial when done.


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OfflinePsilotoad
Monke
Registered: 04/23/22
Posts: 9
Last seen: 2 months, 6 days
Re: Dedikaryotization of Cubensis fruit body clones? [Re: QM33] * 2
    #27781385 - 05/17/22 09:43 AM (1 year, 8 months ago)

Single spore isolation is not as rapid and controllable as dedikayotization. You can buy any eberbach jar and just find a compatible base. Single spore isolation will only yield 1 monokaryon type from the parent and with unknown genetic changes from the parent whereas dedikaryotization can lead to symmetrical recovery of both types allowing the potential of 4 offspring of the same cross from 2 proven parent strains. I performed both and the dedikaryotization is faster and with better results. You can look up SYMMETRICAL RECOVERY OF MONOKARYOTIC COMPONENTS FROM LENTINULA EDODES USING DEDIKARYOTIZATION  R. Ramírez-Carrillo and H. Leal-Lara for further information regarding the advantages of symmetrical dedikaryotization over single spore dilution and other dedikaryotization methods.


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OfflinePsilotoad
Monke
Registered: 04/23/22
Posts: 9
Last seen: 2 months, 6 days
Re: Dedikaryotization of Cubensis fruit body clones? *DELETED* [Re: QM33]
    #28071641 - 11/27/22 02:34 PM (1 year, 1 month ago)

Post deleted by Psilotoad

Reason for deletion: Went too far wasn't constructive


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OfflinePsilotoad
Monke
Registered: 04/23/22
Posts: 9
Last seen: 2 months, 6 days
Re: Dedikaryotization of Cubensis fruit body clones? [Re: Psilotoad]
    #28071646 - 11/27/22 02:35 PM (1 year, 1 month ago)

WhAt iS a 1:1 ratio :rolleyes:


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OfflinePsilotoad
Monke
Registered: 04/23/22
Posts: 9
Last seen: 2 months, 6 days
Re: Dedikaryotization of Cubensis fruit body clones? [Re: wy35]
    #28203660 - 02/25/23 08:25 AM (10 months, 26 days ago)

You could use just water it has no magnesium or phosphorus but the survivability would be lower and the recovery would be much longer. Like Alan said the peptone glucose helps the mycelium grow and recover. Just basically giving it nutes with low magnesium and phosphorus to prevent clamps from forming when submerged in the solution.


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OfflinePsilotoad
Monke
Registered: 04/23/22
Posts: 9
Last seen: 2 months, 6 days
Re: Dedikaryotization of Cubensis fruit body clones? [Re: AyePlus] * 2
    #28203674 - 02/25/23 08:38 AM (10 months, 26 days ago)

When you use multispore each spore that germinates produces monokaryotic hyphae and each successful pairing of monokaryotic strains produces a new dikaryotic strain so when you put down multispore you have many strains of your type. That is why this mentions you want to use an isolated dikaryon. An isolated dikaryon is comprised of 2 genetically distinct types of nuclei one from each parent monokaryon. In di-mon mating only one nuclei type is donated to the monokaryon and this is not a reciprocal exchange. Each dikaryon only has two parent nuclei from their monokaryon parents or mon and di parent but with multispore you are producing many dikaryons. Once you isolate a colony or clone a fruit you have a single dikaryon this dikaryon will be comprised of 2 nuclei one from each parent monokaryon.

Here this includes some more info about nuclei exchange in mon-mon and di-mon pairings

https://www.mdpi.com/2076-2607/9/6/1248/htm


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