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OfflineVisigoth
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Registered: 11/28/01
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agar plate to liquid culture, how do u do it??
    #782514 - 07/28/02 05:48 PM (14 years, 4 months ago)

If I clone a mushroom succesfully on agar plates, is there any way I can start a liquid culture with the colonized agar?? Thanks!!

Vis


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"15 minutes in freezing conditions?!?! You'd become a popsicle before you made that!!!"


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Invisibleangryshroom
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Registered: 12/18/01
Posts: 7,262
Re: agar plate to liquid culture, how do u do it?? [Re: Visigoth]
    #782842 - 07/28/02 09:37 PM (14 years, 4 months ago)

Just wondering, but why would you want to do this? It would be a lot easier to just innoculate the jars with the colonized agar wedges.



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InvisibleChampion des Champignons
long standing member;)

Registered: 07/27/00
Posts: 2,678
Loc: Alba
Re: agar plate to liquid culture, how do u do it?? [Re: angryshroom]
    #782877 - 07/28/02 09:54 PM (14 years, 4 months ago)

...but harder to inoculate a sealed filter patch bag, hmm?
I was wondering the same thing.
I'll probably stick a wedge in a jar of sterile water with some glass shards and maybe a smidgeon of h202 and shake it up to get a mycelial suspension.
Can't think of any other way to do it


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OfflineDarkTranquility
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Registered: 07/06/02
Posts: 72
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Re: agar plate to liquid culture, how do u do it?? [Re: Champion des Champignons]
    #783116 - 07/29/02 01:53 AM (14 years, 4 months ago)

Could you possibly use the blender TEK? ...by Using an entire colonized plate in some water with h2o2 and a little bit of nutrient?.
.....Or what if you cut a piece off of your agar and transfer it to a pyrex tube, shake it with water and h2o2, with something hard inside it to break up the chunk into little bits?


--------------------
"Every picture hods a tale.
Every shade tells of a thousand words.
The artistry of living chaos is
pictured in the poets tears.
The final concept
Is all but a thought away. "


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OfflinestonErollEr1
The Psilocybinsolution
Registered: 05/23/01
Posts: 666
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Re: agar plate to liquid culture, how do u do it?? [Re: DarkTranquility]
    #783150 - 07/29/02 03:27 AM (14 years, 4 months ago)

Wasnt that just what C.d.c said?
Peace
//stonErollEr


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OfflineSuntzu
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Re: agar plate to liquid culture, how do u do it?? [Re: Visigoth]
    #783229 - 07/29/02 05:34 AM (14 years, 4 months ago)

I have used a sterilized cotton-tip swab [PC wood handled ones only!] to swab up some mycelium from the plate, then twisting the swab in some sterilized water to get it dispersed. Repeat.
This gives all sorts of nice little tiny fragments.
Many ways will work well; One tip--try to use the inoculum ASAP.
Good luck;


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InvisibleSumGuy
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Registered: 07/26/01
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Re: agar plate to liquid culture, how do u do it?? [Re: Visigoth]
    #783427 - 07/29/02 07:47 AM (14 years, 4 months ago)

All my friend does is suck up a half a syringe full of sterilized water and squirt it onto the colonized petri, he then scrapes the mycelium off the agar with the syringe needle and sucks up the water back into the syringe and fills the syringe the rest of the way up with water. Works wonders and even if it doesn't look like there's lots of mycelium in the syringe believe me there is and this tek is relatively easy.


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-SumGuy :cool:


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Anonymous

Re: agar plate to liquid culture, how do u do it?? [Re: Visigoth]
    #783468 - 07/29/02 08:14 AM (14 years, 4 months ago)

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Invisiblemycofile
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Registered: 01/19/99
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Re: agar plate to liquid culture, how do u do it?? [Re: ]
    #783521 - 07/29/02 08:58 AM (14 years, 4 months ago)

Much easier, cheaper and reliable is to make a home made eberbach container. See blender usage in niner's tek, or shroomdadi's mycomachine at lycaeum.

BTW, you want to do it because using wedges, a plate innocs about 8 quart jars. Using a blender, that wedge innocs about as many quart jars as you would want. Not to mention you can increase your liquid innoc 10 fold with nutrient broth to innoc a thousand jars. See GG&MM liquid fermentation in the grain chapter. Liquid innoc makes bag culture from agar possible. Bags don't work well with agar, but do stupendously with 200-300 ccs of liquid innoc.


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OfflineShaw
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Registered: 06/28/01
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Re: agar plate to liquid culture, how do u do it?? [Re: ]
    #785814 - 07/30/02 09:17 AM (14 years, 4 months ago)

from my experience, the more liquid you put in, the more chance of contam.


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Anonymous

Re: agar plate to liquid culture, how do u do it?? [Re: Shaw]
    #785922 - 07/30/02 10:19 AM (14 years, 4 months ago)

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OfflineVisigoth
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Registered: 11/28/01
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Re: agar plate to liquid culture, how do u do it?? [Re: ]
    #786772 - 07/30/02 06:21 PM (14 years, 4 months ago)

Couldn't you transfer an agar wedge into a jar of honeywater or a solution made with powdered malt extract or something?? I thought that would be better than plain old water because it would be nutritious to the mycelium and promote growth, no??

Vis


--------------------
"15 minutes in freezing conditions?!?! You'd become a popsicle before you made that!!!"


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OfflineSuntzu
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Re: agar plate to liquid culture, how do u do it?? [Re: Visigoth]
    #787560 - 07/31/02 05:33 AM (14 years, 4 months ago)

It depends somewhat on what your goal is. It seems quite sufficient to just fragment the mycelium in liquid, giving many inoculation points. For some species it helps to let the fragmented cells regenerate and branch before inoculating--this would be where some nutrient is helpful.
The downside to using nutrient solution is that it can also breed contaminants. This is going to vary from person to person. Something like dH2O is less likely to be a problem, assuming tools are clean, etc.
People have different success stories with a variety of techniques. Personally, I like using swabbed up dH2O inoculum because of its simplicity. Test tube, water, swab!
Good luck


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