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Offlinegt40
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Anybody here have contam in petri dishes before inoculation?
    #18922490 - 10/02/13 05:14 PM (10 years, 3 months ago)

:crazy:
It happened for a few times. I took my dishes from PC and put it in a plastic bag. One day later when I want to start inoculation, I saw contam on agar in all petri dishes. I have no problems with sterilization before.


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OfflineHabadashery
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Re: Anybody here have contam in petri dishes before inoculation? [Re: gt40]
    #18922570 - 10/02/13 05:30 PM (10 years, 3 months ago)

ONE day later?  Oye!

Don't know.  Even if you didn't PC it long enough or hot enough, I would think it would take more than ONE day to contam.


Hmm.


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InvisibleSpitballJedi
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Re: Anybody here have contam in petri dishes before inoculation? [Re: gt40]
    #18922725 - 10/02/13 06:08 PM (10 years, 3 months ago)

I presume you are using glass dishes since you said you took petri dishes from the PC.

On a highly nutritive media, molds can grow pretty quick, but one day is fast.

What tek are you following?

I don't trust the glass ones because I don't feel like I can keep them sterile from PC to storage.

The bags should be brand new ziplocks.

Quote:

gt40 said:
:crazy:
I have no problems with sterilization before.




When we do things imperfect, sometimes we still get great results, sometimes for a great while even. Only when we run in to problems do we realize we can make improvements to get consistent great results. The devil is in the details.


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OfflineRogerRabbitM
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Re: Anybody here have contam in petri dishes before inoculation? [Re: gt40]
    #18923116 - 10/02/13 07:52 PM (10 years, 3 months ago)

You're supposed to sterilize the agar in a bottle and then pour it later.  If you're using glass petri dishes, just stick them in the PC empty while you sterilize the agar.
RR


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Offlinerumfor69
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Re: Anybody here have contam in petri dishes before inoculation? [Re: RogerRabbit]
    #18923428 - 10/02/13 09:12 PM (10 years, 3 months ago)

When you pour agar how would you sterilize plastic petri dishes before hand?

Or do you just buy them pre-sterilized?


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OfflinePussyFart
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Re: Anybody here have contam in petri dishes before inoculation? [Re: rumfor69]
    #18923435 - 10/02/13 09:13 PM (10 years, 3 months ago)

Quote:

rumfor69 said:
you just buy them pre-sterilized




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Offlinerumfor69
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Re: Anybody here have contam in petri dishes before inoculation? [Re: PussyFart]
    #18923450 - 10/02/13 09:16 PM (10 years, 3 months ago)

Quote:

Notahacker420 said:
Quote:

rumfor69 said:
you just buy them pre-sterilized








Thats what i figured i just had to ask LOL


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Offlinegt40
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Re: Anybody here have contam in petri dishes before inoculation? [Re: rumfor69]
    #18924357 - 10/03/13 02:55 AM (10 years, 3 months ago)

Wow, thanks for fast responde! For sure I have used glass dishes. Every time A LOT of condensation inside after PC. Maybe it initiates contam, although I put them one by one. I think I will try to prepare the agar in the bottle separately next time.


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OfflineHabadashery
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Re: Anybody here have contam in petri dishes before inoculation? [Re: gt40]
    #18924920 - 10/03/13 08:19 AM (10 years, 3 months ago)

And really, using 1/4 or half pint jars with filter lids, is way more user friendly imo.  Most of the petri dishes I tried to make in the past failed, just couldn't keep it together.  For those of us without the steady hand, I would suggest agar in a jar.  Haven't had but 1 fail in a jar so far, outta 20+

And its a great thing to do with those jars you used for PF tek, which you probably aint never gonna do again!


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InvisibleFred Teddy
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Re: Anybody here have contam in petri dishes before inoculation? [Re: Habadashery]
    #18925149 - 10/03/13 09:48 AM (10 years, 3 months ago)

Quote:

Habadashery said:
And really, using 1/4 or half pint jars with filter lids, is way more user friendly imo.  Most of the petri dishes I tried to make in the past failed, just couldn't keep it together.  For those of us without the steady hand, I would suggest agar in a jar.  Haven't had but 1 fail in a jar so far, outta 20+

...




Petri's "vent" thats why you're supposed to seal it up with parafilm.

I do agar both ways but I like the no-pour agar in 1/4 pt jars. I use a lid seal side down with a large 3/4" hole in it to help the condensation escape. Seal side down because I always seemed to get contaminants around the edges. That stopped when I started using seal side down. Use whatever filter you want. I use tyvek held down with a ring magnet. Cover with foil and PC. Remove from PC, remove the foil, and let the jars cool. You will get little or no condensation. To store or keep from drying out, I put a piece of plastic bag over the filter and put the magnet back on. From here, once you determine a clean sample, you can continue to do agar work, or let go to 100% and add an LC or sterilized water for syringes, or do a sterilized grain to agar jar (tricky though, too much exposure). I like the LC approach, I can do more grain jars that way than just using the agar as wedges, and I know its clean. I know this is not "traditional" but a jar of myc has a lot of versatility imo.

Peace,
FT


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OfflineRauhfasertapete
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Re: Anybody here have contam in petri dishes before inoculation? [Re: gt40]
    #18925275 - 10/03/13 10:31 AM (10 years, 3 months ago)

Quote:

gt40 said:
:crazy:
One day later when I want to start inoculation, I saw contam on agar in all petri dishes.





I don´t think you can get visible contamination in one day. But I once had something that looked similar. I boiled potato peels together with baker´s yeast and fructose, and then let the starchy cooking water drip through a coffee filter. The resulting liquid was quite homogenous, and I used it for my agar which I then sterilised in the petri dishes. When it was solid again after PCing, I noticed that some kind of little spots (about a mm in diameter) have formed in the agar, especially on the bottom. They seem to have formed from starch or other small particles.

It still worked well, actually some of the finest agar I evervmade. It just didn´t look nice.


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Invisible36fuckin5
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Re: Anybody here have contam in petri dishes before inoculation? [Re: Rauhfasertapete]
    #18925644 - 10/03/13 12:25 PM (10 years, 3 months ago)

Quote:

Rauhfasertapete said:
Quote:

gt40 said:
:crazy:
One day later when I want to start inoculation, I saw contam on agar in all petri dishes.





I don´t think you can get visible contamination in one day.




Of course you can. That's just silly.

OP, wrap your empty petris in foil and PC them, then pour them in a SAB or in front of a flowhood.


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