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 kaliman
Researcher
Registered: 02/12/08
Posts: 44
Last seen: 4 months, 18 days
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 Innoculating LC with agar, tissue, spores or LC
#9686517 - 01/26/09 11:06 PM (3 years, 3 months ago) |
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I was wondering that there is not (I have not seen) a complete guide on different ways to inoculate a LC.
There is plenty of info about spores into water into siringes.
Good about LC to LC
There is way less about live tissue.
There is very little about agar.
Other thing
I have 3 petri dishes of Agaricus bitorquis, Pleurotus djamor, and Stropharia rugosoanulata.
They seems to be kind of not totally white. Kind of light covering.
It seems difficult to cut w a scapel a clean cap of mycelium and I think I will have some part of agar with it.
I think that ussually agar should not go with mycelium and I have not seen anybody doing it. I suposse that its not used because the idea is to separate contams and if you put agar you are possibly keeping some?
What if I put a little of agar with the mycellium?
And going back to the original point:
Which would be the best way to inoculate a LC with
a) Live tissue
b) Agar
I mean, how much, procedures, Glovebox, bottles with vials, etc???
Any opinion??
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Kalimantan is the Paradise
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 shymanta
Mad Scientist
Registered: 01/27/05
Posts: 904
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Re: Innoculating LC with agar, tissue, spores or LC [Re: kaliman]
#9686703 - 01/27/09 12:03 AM (3 years, 3 months ago) |
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You can use a syringe to squirt a little water onto the surface of the colonized agar (it must be clean of contamination) and use the needle to rough up the mycelium a bit. Then draw the water back into the syringe capturing as much mycelium as possible. Do this in a glove box or in front of a flow hood.
Then you can inoculate the LC.
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kaliman
Researcher
Registered: 02/12/08
Posts: 44
Last seen: 4 months, 18 days
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Re: Inoculating LC with agar, tissue, spores or LC [Re: shymanta]
#9687980 - 01/27/09 10:02 AM (3 years, 3 months ago) |
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Thank you Shymanta
I know that is a procedure, actually the most quoted of all for going agar to LC.
I also see Stamets (Growing Gourmet... chapter 15) using a quite big square cutout of mycelium to inoculate a Eberbach stirrer.
So we have two procedures here, wedges and water on agar.
I think water on agar maybe is very appropriate in some situations because you can use it with syringes. But some mycelium are harder to dissolve and some are easier, and how much agar will be dissolved in the solution? Also maybe the soft cotton like edges maybe will dissolve before the strong rhizomorphic parts?
I have made some syringes by sucking a piece of live tissue into a syringe and then using it as my LC. I have done it while cloning mushrooms from the shop and sometimes wild species. It works. Somebody else invented the Tech, not me.
There are other teks however.
I guess your vote as the best tech is puring sterilized water into the agar to suck it with syringe. Would you mind to say why??
Thanks
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Get a happy day today
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