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InvisibleJoshua
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Registered: 10/27/98
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Agar plate info?
    #443171 - 10/31/01 03:09 PM (15 years, 9 months ago)

I would like to transfer some mycelium that I have growing directly to an agar plate. I have never used the agar method before and am looking for some pointers. I would like to use the agar method as I have innoculated a brown rice cake with some old spores, the growth is very slow inside my jar and contamination is starting to set in. I would like to isolate my good mycelium and continue with it. The spores I have are from a very large cubensis strain.

I plan on plating the agar and transfering active mycelium to the plate. I will incubate at an appropriate temp until I get some rhizomorphic growth. It is at this point that I'm lost. Once I get mycelial growth on my plate how should I procede with innoculation of my substrate?

Any pointers would be helpful, e.g. agar recipes, incubation time/temp, etc.

Thank you,
Joshua


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Offlinefelixhigh
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Re: Agar plate info? [Re: Joshua]
    #444166 - 11/01/01 02:27 AM (15 years, 9 months ago)

buy the mushroom cultivator. i think everybody who wants to cultivate shrrmmz should have this book around...


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OfflineElektrolurch
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Re: Agar plate info? [Re: Joshua]
    #444372 - 11/01/01 06:07 AM (15 years, 9 months ago)

Like felixhigh wrote, for such questions you should get "The Mushroom cultivator". But I can still give you some hints, so you can work until you get it...

After you selected some good looking mycellium on agar, just let it grow on agar again until 3/4 of the dish arecovered.
Cook some rye or any kind of seed substrate. Cut with a sterile knife, etc. some triangles (wedges) of agar with mycellium on it. After use this to inoculate the above substrate (rye etc.)

The easiest agar reciept is MEA:

20 g malt extract
20 g agar-agar
4 g pepton (optional)
4 g yeast extract (optional)
fill up to 1 litre with water

Have luck,

Elektrolurch


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The doll's house, darkness, old perfume
And fairy stories held me high on
Clouds of sunlight floating by.", Pink Floyd '67


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InvisibleNDK
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Registered: 07/13/01
Posts: 186
Re: Agar plate info? [Re: Joshua]
    #444393 - 11/01/01 07:00 AM (15 years, 9 months ago)

You'll make your life a lot easier if you use peroxidated agar. When you made up the agar and sterilised it, you let it cool so it is only warm to the touch. You then squirt in some 3% H2O2. I use 10ml per litre. (at litre is a lot of agar BTW!). Then when you transfer your mycelium you are a lot less likely to pick up contaminants.

I'd make up about 5 plates and take 5 samples of mycelium to make sure you get what you want. Bear in mind that the mycelium you transfer will stay as it is i.e. if it is cottony it will not turn into rhizomorphic, so try to find the best stuff. Use a flamed scalpel for the transfer. For extra sterility you can dip the chunk of mycelium into neat peroxide before the transfer. This will kill most contaminants that might already be on the mycelium but can slow/kill some weak mycelium on rare occasions.

For general agar tips, see Fat Guy's Agar Tek in the Grow or Find section.


Edited by NDK (11/02/01 12:02 PM)


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OfflineAnnoA
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Re: Agar plate info? [Re: NDK]
    #444434 - 11/01/01 08:08 AM (15 years, 9 months ago)



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OfflineTreeofLife
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Re: Agar plate info? [Re: Anno]
    #444826 - 11/02/01 04:17 PM (15 years, 9 months ago)

where do you buy agar plates?>


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The only thing you have to fear is, fear itself...


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Invisibledimitri211
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Registered: 04/27/01
Posts: 2,248
Re: Agar plate info? [Re: TreeofLife]
    #444972 - 11/02/01 06:36 PM (15 years, 9 months ago)

myco supply.com
fungi.com
drbluethumbs for 3.50 a sleeve


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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