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InvisibleNillion
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Registered: 04/14/22
Posts: 1,000
Loc: Terra Firma
Re: Dedikaryotization of Cubensis fruit body clones? [Re: cheor] * 1
    #27733880 - 04/14/22 05:36 AM (1 year, 9 months ago)
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I'd try a variant of Leal-Lara and Egar-Hummel 1982.

They had some challenges regarding the ratios.
The method employs magnesium and phosphorus deficient media and carefully controlled volumes to prevent overcrowding so that the torn hyphae and their nuclei can recover and become Neohaplonts. If the density is too high they reconnect.

The Peptone P method takes advantage of the high glycine content and the use of simple media made of anhydrous glucose and peptone P or glycine prevents caramelization and or sediment from forming in the media.

This method for production of monokaryonic neohaplonts using dedikaryotization has been employed for decades.

I was able to obtain the 1982 paper for further consideration and have attached it to this post.


Edited by Nillion (04/14/22 05:41 AM)


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InvisibleNillion
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Registered: 04/14/22
Posts: 1,000
Loc: Terra Firma
Re: Dedikaryotization of Cubensis fruit body clones? [Re: wy35]
    #28599977 - 12/27/23 03:56 PM (1 month, 5 hours ago)

Quote:

wy35 said:
What makes the peptone and glucose solution more dedikaryotizing than just water?



It is actually slightly deficient in nutrients, which stunts the growth of the mycelia and makes it weaker, leading to increased production of neohaplonts, which can appear and recombine rapidly and thus be difficult to recover in some situations. A key part of the method is also culture density, by slowing growth and avoiding excess density it becomes easier to isolate the neohaplonts.

Also, the blender is nothing more than an agitation method, hand agitation or another method of stirring can work just as well to create the sheer forces needed for this, the blender is just faster. Any blender you can keep sterile works just fine for this.


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InvisibleNillion
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Registered: 04/14/22
Posts: 1,000
Loc: Terra Firma
Re: Dedikaryotization of Cubensis fruit body clones? [Re: fahtster] * 1
    #28599983 - 12/27/23 04:01 PM (1 month, 5 hours ago)

Quote:

fahtster said:
Not sure if you guys have seen This thread but I seemingly have crossed compatible clones by blending pulverized myc water of two clones and inoculating qts directly with the mixed




Yeah, one doesn't need to isolate neohaplonts to use dedikaryotization for breeding, one can just agitate mixed cultures prior to inoculation and screen the results for the desired phenotypes. It's quite handy actually.


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