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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Agar mistake - new LC method? ::update::
    #6893182 -

On my first attempt at making agar plates, I used half pints and homemade PDA. Upon trying to inoculate through the taped up hole in the jar lid, I found my needle was too short, so I just went ahead and shot some solution in there anyway.

Now, I am not the smoothest with the plunger, and quite a bit of solution ended up in the dish (it was an old syringe, that I didn't believe was viable anyway...). I sloshed the solution around in the agar jar, not quite realizing that this kind of defeats the point of the agar dish.

Within the next couple of days, I ended up with a milky solution floating around on top of the agar. Hmmm, I thought...

I prepared some sterile dextrose water and filled a used, sterilized syringe. I stuck the needle through the hole, and sucked up the milky solution on top of the agar. Within the next few days, I had big, visible strands of mycelium floating in the syringe.

I left it for a week or so, out of circumstance, and used it on some BRF jars yesterday. No growth yet, but I'm sure that's to be expected. I'll update when there is growth.

Not to say this takes the place of any other LC technique, but I found it pretty fast and easy to do, at least by accident.

Any thoughts, suggestions, questions, worries, anything?


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“I thought naming myself ‘ReoSpeedwagon153’ on a forum was a funny idea in 2006.”

Edited by ReoSpeedwagon153 (05/14/07 11:05 AM)

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Re: Agar mistake - new LC method? [Re: ReoSpeedwagon153]
    #6893329 -

Quote:
I ended up with a milky solution floating around on top of the agar


That would be bacteria. A contaminant.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Agar mistake - new LC method? [Re: RogerRabbit]
    #6893552 -

I don't know though...

I still have the agar jar, which I've kept in kind of a cold spot. It's starting to show rhizomorphic growth on the surface of the agar, and on the side of the jar where it made contact with the solution when it was tipped to suck up the water.

It's clearly mycelium, and not bacteria which should be colonizing the agar if that's what the milky solution was.

My description of it as 'milky' may not be completely accurate. I really wouldn't know how to even start giving it an accurate description...

I guess I'll just see what happens.


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“I thought naming myself ‘ReoSpeedwagon153’ on a forum was a funny idea in 2006.”

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Re: Agar mistake - new LC method? [Re: ReoSpeedwagon153]
    #6893800 -

Here's what I was referring to as 'milky'. Sometimes I use the word 'greasy' as well. The top right section has bacteria contamination. The top left section has satellite colonies, probably molds. Only the bottom section is good. This is a clone from a wild fruit, but it shows some of the stuff to look for when doing agar work. Good luck with your projects.
RR



--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Agar mistake - new LC method? [Re: RogerRabbit]
    #6913684 -

Not sure about the moldy looking ones, but it definetely isn't the milky bacteria. I think it was just some kind of mycelium slurry from having so much extra spore solution in with the agar.

Update:

Five days from inocculation, one jar shows fuzzy growth at two of the inoc points. Other jars still nothing. I'll keep an eye on it.
_____________________________________________________________________
Oh yeah, and a question about my agar work: My needle doesn't reach to the agar (half-pint jar with a hole for inocculation), so I am forced to squirt a little solution onto it instead of into it. This leaves solution floating around on top and inocculates the entire surface of the agar, instead of just one point from which mycelium would grow. Is this even a problem?


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“I thought naming myself ‘ReoSpeedwagon153’ on a forum was a funny idea in 2006.”

Edited by ReoSpeedwagon153 (05/14/07 11:13 AM)

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Re: Agar mistake - new LC method? ::update:: [Re: ReoSpeedwagon153]
    #7068925 -

I'm only putting the bump on here as an update, in case for some reason someone comes across this thread and wants to know if my attempt was successful or not. I hope that is 'truly relevant.'

The jars I made eventually colonized and have been cased. I see one pin and many lovely hyphal knots ready to light up.

The agar dish grew over and formed many clusters of primoridia, at which it has now been stalled for a week or so.

The syringe was used again to inoculate my first attempt at WBS jars and they were attacked by a green mold.


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“I thought naming myself ‘ReoSpeedwagon153’ on a forum was a funny idea in 2006.”

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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