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WBS Cloning

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Posted by: Mycota Mar 02 03, 03:54 AM GMT

WBS CLONING

Prep & PC several pint WBS Jars (with internal Tyvek filter & allow to cool to room temp.

Select & harvest a healthy large fresh mushroom specimen (under the cleanest possible conditions).

Give the stem of the mushroom a good dousing with Hydrogen Peroxide

With sterile instruments (knife, scalpel, exacto blade, tweezes, cuticle scissors or whatever) under the cleanest possible conditions, open the stem & remove several pieces of interior flesh & drop several pieces of it into each pint jar & give them a mild shake (to disperse the mushroom pieces into the WBS).

Cap & incubate as you would a normal WBS jar. Colonization will be slower than normal WBS spore -or- G2G transfers, as there are only a few points of contact between the small pieces of mushroom flesh & the WBS, in the jar.

Once colonized, you have created an "isolate".........aka........"clone" of the specimen.

To propagate that further, simply G2G that colonized WBS into other larger (quart) WBS jars, allow them to colonize & spawn as usual, into you favorite substrate.

Mycota (aka 6T)

Posted by: nobody Mar 02 03, 05:42 AM GMT

Interesting technique. Would you recommend washing the cuttings in H2O2? Have you thought pur�eing the tissue to get more points of inoculation.

Posted by: dcyans Mar 02 03, 09:01 AM GMT

I've took a clean interior piece and in the h2o2 filled cap i cut and diced the piece. Then place on agar,i like to break the surface of the agar and push the piece into the agar. three of four attemps were successful, one was too small a piece i think and it just dried up. itried this after reading a post by FM

Thanks again FM

for this one

Posted by: Mycota Mar 02 03, 12:53 PM GMT

QUOTE (nobody @ Mar 02 03, 10:42 AM GMT)

Interesting technique. Would you recomend washing the cuttings in H2O2? Have you thought pur�eing the tissue to get more points of innoculation.

You can wash any tissue in mild h202. You can puree, if you have the means to & keep it clean. But, adding a step, adds that much more potential to intro a contam.

This method isn't fast, but requires nothing fancy, no agar, plates & works about 60% of the time.

That is why I suggest using pints or even 1/2 pints, because they will colonize quicker & doing several (like 5 or 6) at a time. All it takes is one good one, to go & your off and running via G2G.

Mycota

Posted by: glasshoppa Mar 02 03, 01:07 PM GMT

What is the advantage of cloning over multi-spore inoculation?

Posted by: Mycota Mar 02 03, 01:18 PM GMT

Multi/spore innoc is the whole mix, good, bad, weak, strong, big & small, all in one shot.

Advantage is selective breeding, as you get the genetics of a big healthy shroom, instead of a gang bang mix.

Like breed a big dog with one big stud. You get BIG puppies.

Let a big bitch dog breed with 20 assorted street muts, what do you get?

See the logic?

Mycota

Posted by: eatyualive Mar 02 03, 01:36 PM GMT

a foaf tried this method but it failed bc of rye berry! but it does work!

might i add, nice logic!

Posted by: glasshoppa Mar 02 03, 01:46 PM GMT

OK Mycota, I see what you're saying. Be patient with me here as I try to understand...

When I inoculate with spores from a single print, don't I already have all the genetics of that one mushroom in the spores that all came from the same cap? Are you saying that their is genetic variation in all the spores dropped from a single mushroom?

Posted by: OneDiaDem Mar 02 03, 01:52 PM GMT

LOL, think of them as sperm, some make it, some dont, some are weak, and some just, well, you get the idea.

Posted by: Mycota Mar 02 03, 02:37 PM GMT

QUOTE (OneDiaDem @ Mar 02 03, 06:52 PM GMT)

LOL, think of them as sperm, some make it, some dont, some are weak, and some just, well, you get the idea.

LOL, think of a multi/spore innoc like "sperm" from 100,000 men, rather than one select stud (as in a clone).

Mycota

Posted by: Zoom Mar 02 03, 04:55 PM GMT

That's..........Really kinky.

Posted by: nobody Mar 02 03, 05:10 PM GMT

The genetics in fungal reproduction seems to be a bit more complicated then that of animals.

QUOTE

"The number of mating classes is somewhat greater then in bipolar forms, since four types arise from spores of a single basidiocarp. Obviously thesethyes, numbering in the hunders in both bipolar and tetrapolar species, cannob be designated as sexes! (Scagel, et. al.., 1967, p.69)

I'm nobody.

Posted by: glasshoppa Mar 02 03, 11:44 PM GMT

QUOTE (OneDiaDem @ Mar 02 03, 10:52 AM GMT)

LOL, think of them as sperm, some make it, some dont, some are weak, and some just, well, you get the idea.

Um, thanks. I'll never think of inoculation quite the same way again...

Shroom Glossary