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OfflineUnderNose
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Contaminated petri dishes.
    #6666905 - 03/14/07 12:31 AM (14 years, 8 months ago)

I was not sure if this should be in contamination forum of here but anyway
Best to start from the start.

I received a few prints for some wild Pan Australians, This would be my first attempt at Panaeolus mushrooms.

After some advice I thought I would get some petri dishes & PDA.
I was told that this could increase the chance of success by isolating good sections of growth from the dish & also from possible contaminants that the "wild" print may have.

I also want to be able to clone other shrooms & keep samples of good ones in the fridge.

I ordered some dishes & agar but  I decided to also see what happened if I just dumped one of the prints into a LC.

To my surprise the LC looked like it was growing nicely with no cloudiness just very fine mycelium that looked quite different than the cubensis mycelium that I am used to seeing.
It progressed quickly & about 5-7 days after putting the spores in I decided to start some grain jars, At this stage the agar & other things had arrived.

Here are the jars of WBS after a week or two. almost ready to spawn I think


Now the other day I decided to test out the petri dishes.
Agar was prepared & poured in the most sterile manner I could.

I got the pan LC that still looks even today to be good & with a 3ml syringe I placed one drop of the LC onto 8 dishes. I also did two petri dishes  with a cubensis strain LC.
Right after doing this I thought to myself that it might have been better to use prints instead.


Three days later all the Pan petri dishes were growing this.
 

It smelled quite bad when I took this photo even with the petri dish next to a open window. After the photo all 8 dishes were discarded.
The cubensis ones looks good, Little white fluff starting


So why would the LC & jars "look" like they are colonizing nicely but the petri dishes are not.?
Anyone know what this crap growing on the dish is.?
Is it possible the grain jars are contaminated also.?
Is it better to start a petri dish with a print or LC.?

This is my first attempt at agar & pans so some help would be great.
Thanks:grin:


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:dna::dna:


Edited by UnderNose (03/14/07 12:49 AM)


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Invisiblefastfred
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Re: Contaminated petri dishes. [Re: UnderNose]
    #6667665 - 03/14/07 06:21 AM (14 years, 8 months ago)

I would say that the WBS looks contaminated. Hard to say with what. It could be myc coated with yeast contamination.

The petri dish is a contaminate for sure. You can tell by the rounded margins. The wrinkles in the center are another sign. I would guess yeast, but it could also be bacteria.

Your best bet is to isolate on agar or do multiple cultures using just a quick scrape of spores each.


-FF


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OfflineRogerRabbitM
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Re: Contaminated petri dishes. [Re: fastfred]
    #6668434 - 03/14/07 01:28 PM (14 years, 8 months ago)

Definitely contaminated. This shows why it's important to culture onto agar and not LC, where you can't see the contaminants until much later, wasting valuable time. I'll move this to the contaminanation forum for you. That is where it should be.
RR


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OfflineUnderNose
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Re: Contaminated petri dishes. [Re: RogerRabbit]
    #6671189 - 03/15/07 01:55 AM (14 years, 8 months ago)

Thanks for moving my post roger, I knew I had it in the wrong place.

Thanks also FF.

Yes I know my petri dished are contaminated, They were chucked in the bin just after that photo.
So do you think the grain jars that were made from the same LC will also be contaminated.
It's a little hard for me to tell as I have never seen Pan myc

Am going to start some more but this time I will use spores not LC


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OfflineRogerRabbitM
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Re: Contaminated petri dishes. [Re: UnderNose]
    #6672059 - 03/15/07 11:56 AM (14 years, 8 months ago)

Pan mycelium is linear and thin, not rhizomorphic. Unfortunately, there was no mycelium visible in that petri dish to transfer away from the bacteria. I'm sure anything you made from that lc is also contaminated. Next time, forget the lc and start your spores on agar.
RR


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