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Invisiblemorty422
Wuba-luba-dub-dub!
I'm a teapot


Registered: 07/06/16
Posts: 988
On the subject of cloning...
    #23595434 - 08/30/16 08:55 PM (7 years, 4 months ago)

Quote:

FrankHorrigan said:
This is part of my series on how I get things done. I hope it helps!

The easiest way to get a good set of genetics is to clone an ideal looking fruit or cluster from a multispore grow. This process is very simple, and the way I do it is essentially the same way that RR does it.

Some people will try to clone into LC or using a needle, etc. This will almost always fail. Make sure to use agar. Since fruiting is done in open air, you can always assume the fruit is covered in contaminants. This is doubly true for cloning wild mushrooms.

Even though you'll be taking a cut of virgin mycelium that has never been exposed to open air, it is very common to need to transfer your mycelium away from contaminants on the first dish.

Since a flowhood can potentially blow contaminants around from the fruit you are cloning, I recommend working in a still air box.


Start by wearing tyvek sleeves and disposable gloves. Make sure to work quickly and carefully inside the still air box. Use good sterile technique.

Flame sterilize your scalpel and set it down in the still air box where it won't be disturbed. I place it on my receiving petri dish. Do not set it on the floor of your SAB.

Take the mushroom (this is a golden oyster) and pinch it at the base. Rip it down the middle. Do not cut into the stem with your scalpel or you will force contaminants down into it.



Holding part of the fruit with one hand, use your scalpel to gently scrape at the center of the stem, near the base. Try to get a small piece on to the blade. As RR says, a scraping motion will be easier than trying to slice a specific piece.


Once you have a small piece of tissue on the end of your scalpel, transfer it to the agar. It may take a bit of effort to get it off the blade :shrug:


Take as many pieces of tissue as you want to clone. You can flame sterilize between each cut, or just flame sterilize between each petri dish or fruit. I use 3 section dishes so I flame sterilize every three cuts.

That's all there is to it. Wrap the petri dish and let it colonize. Transfer healthy mycelium away from any contaminants as soon as they appear!

Some fruits will give you an isolate right off the bat, but a lot of mushrooms are actually composites of multiple genetics. However, I have seen TCs say that the original clone sectors perform better together than as an isolate of each. So it's up to you if you decide to isolate the clone further :thumbup:


:cheers:





In the bold text of Frank's post above he states that some TC's say the original clone sectors perform better TOGETHER than as an isolate of each...

This might be hard for me to ask - being new to AGAR I may possibly be using the wrong terms. Bare with me, correct me as I go and let me know if I am on the right track with this ideology!

1. The cloned tissue of the best fruit is transferred to agar.

2. The cloned tissue is cleaned up further by transferring a section of clean growth that show no signs of bacteria.

3. The next dish of agar is very clean (if not step 2 can be repeated once more)and can be colonized fully to be dropped into multiple masters or possibly just one jar (such as the tiger drop).

^^^^Let me know if any of this was wrong, even in the slightest- as the next portion of my post and my actual question sort of revolves around the top being true.

That final plate (before the drop into grain, or LC, or whatever) could possibly look like this:



or this:



My questions regarding this are:

A: If you are trying to clean up a fruits tissue to clone it, are you also separating and isolating 'away' some of the genetics as you go?

B: If you are "tearing up that fruits genetics" are you now diminishing the chance that your cloned grow will grow that type of fruits characteristics?

C: (I think this is where Frank's quote comes into play...)
Is the monoculture what we are after? If question A is true, is it best to isolate sectors as little as possible to keep that fruits genetic code present in your grow?

D: Is a good goal to continue with multiple grows of the same mycelium and continue to clone fruits off the same genetics you cleaned up before? Is there anything that could even come of that-that is worth the time?

E: Is all this shit nebulous and I just need to grab some good-clean rhyzo myc and roll that bitch out?

Thanks Cult!


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