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Registered: 05/13/03
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innoculating with tissue?
    #1544390 - 05/13/03 08:19 AM (14 years, 17 days ago)

I've got a big old pile of p. Tamp., and I want to clone them... Now, i know you can put a sterile slice in growth medium, then use that to innoc. jars/bags, but it seems to me you ought to be able to use the tissue slice directly onto the growing substrate. Am i wrong? (i should think so...)

If i do need to put the tissue in a liquid "broth" first, can anyone recommend mix that works -- ive seen a lot of teks, ranging from maple syrup/water to complicated agar/dextrose/malt/yeast madness?

TIA everone, and keep up the good work



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OfflineAnnoA
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Re: inoculating with tissue? [Re: sideshow]
    #1544435 - 05/13/03 08:55 AM (14 years, 17 days ago)

* inoculating

> you ought to be able to use the tissue slice directly onto the growing substrate

true.


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Invisiblemicro
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Re: inoculating with tissue? [Re: Anno]
    #1544492 - 05/13/03 09:36 AM (14 years, 17 days ago)

You could use a sterile piece of a fruitbody in the jars (after they cool) but it would take longer to colonize because it will be harder to spread the hyphal cells.

Any liquid medium suitable for fungi should work; it will even grow a bit in dextrose water.  I would use agar first, IMO, to isolate a known-to-be clean sample and then throw that in the water.

If you need any help, tell me -- I am great at ghetto rigging media recipies.  Malt/Dextrose/Yeast extract sounds good, though -- the yeast is for vitamins like biotin so it can be good to add in -- makes for healthier growth.  I go so far as to even throw in a non-essential amino acid mix :wink:

Just if you make liquid media, remember 2 things:

1.) STERILE (of course.)  You could always put in a tight-fitting septum plug (like on the blood vials) to ensure sterility.

2.) You're probably going to want to break up the mycelium up afterwards.  You'll want to put something like broken glass or metal jacks in it (I use 1mm glass beads if you happened to work in a lab or one at school they should have some.)  Anything that will sterilize ok.  Large ~16g needles also help to draw it up.

--
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Re: inoculating with tissue? [Re: micro]
    #1544541 - 05/13/03 10:34 AM (14 years, 17 days ago)

Thanks for that micro - just a couple of qs:

I'm growing for sclerotia, so time isnt a problem - if i chuck sterile slices of truffle in a jar of medium, will that (eventually) form sclerotia if i just leave it? And what medium would you recommend - i don't have acces to a lot of gear (or ??), so i was thinking white rice + water (recommended for sclerotia formation). Would adding dextrose/yeast to that help?




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Invisiblemicro
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Re: inoculating with tissue? [Re: sideshow]
    #1544705 - 05/13/03 11:44 AM (14 years, 17 days ago)

Not if you're trying to form sclerotia; you want a more simplistic medium.

Are you trying to grow truffles? Noone has done this as of yet -- they grow way underground and their spores are spread through mole feces. Some PhD did patent some way to innoculate a forest in Oregon, though.

You might be able to grow the mycelium, but I seriously doubt you could get it to fruit. Sorry, I don't know if it forms sclerotia.

Oh -- FYI -- you'd need a fruitbody less than 48 hrs old.

--
Micro


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Edited by micro (05/13/03 11:45 AM)


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Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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