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OfflineTheCyndicate
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What a contaminated plate!!
    #24035192 - 01/24/17 05:44 AM (7 years, 6 days ago)

So on 1/22 Inoculated 20 dishes with a MS syringe
I used a sterile swab for each dish that was wrapped in its own tin foil then placed in a pint jar with a lid with polyfill. And PC at 15 PSI for 20 min along with the same agar media that was used with for these dishes . All dishes where infected with bacterial contamination with in 3 days .

In my opinion it was the spore syringe that was contaminated. And not my sterile technique. Given the contamination occurred only where I seabed the agar . I even did a smiley face one and it is contaminated in the shape of the smiley face.



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OfflineHaywire
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Re: What a contaminated plate!! [Re: TheCyndicate]
    #24035209 - 01/24/17 06:01 AM (7 years, 6 days ago)

There's two possibilities in my opinion.
It could indeed be the spore syringe.
but also the swabs.

liquids are self sterilizing. if you bring them to 121°C they will kill everything that's dissolved in it.

a closed pintjar with filterpatch, filled with air, will take a lot longer to fill with steam. 20 minutes is perhaps too short for the pintjar to sterilize.


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OfflineTheCyndicate
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Re: What a contaminated plate!! [Re: Haywire]
    #24035218 - 01/24/17 06:12 AM (7 years, 6 days ago)

Quote:

Haywire said:
There's two possibilities in my opinion.
It could indeed be the spore syringe.
but also the swabs.

liquids are self sterilizing. if you bring them to 121°C they will kill everything that's dissolved in it.

a closed pintjar with filterpatch, filled with air, will take a lot longer to fill with steam. 20 minutes is perhaps too short for the pintjar to sterilize.




Good thoughts. But for the entire batch to be infected with the same bacteria when a new swab was unwrapped and used for each dish is kinda tell tell IMO . I do appreciate your thoughts and will definitely take them . And I will increase my PC time to compensate. Thanks for the insight.


Edited by TheCyndicate (01/24/17 06:41 AM)


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OfflineHaywire
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Re: What a contaminated plate!! [Re: TheCyndicate]
    #24035232 - 01/24/17 06:24 AM (7 years, 6 days ago)

Quote:

TheCyndicate said:

new swab was unwrapped and used for each dish is kinda tell tell IMO .




I had indeed not taken that into account!


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OfflineTheCyndicate
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Re: What a contaminated plate!! [Re: Haywire]
    #24036593 - 01/24/17 04:51 PM (7 years, 6 days ago)

It's the only thing I can think of. If anyone else can think of anything let me know.


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OfflineOrbit
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Re: What a contaminated plate!! [Re: TheCyndicate]
    #24036644 - 01/24/17 05:09 PM (7 years, 6 days ago)

Most likely the sryinge. But why not just put a drop on the plate and streak with a sterilized a loup. You can just use a piece of wire after flaming.


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OfflineTheCyndicate
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Re: What a contaminated plate!! [Re: Orbit]
    #24036878 - 01/24/17 06:23 PM (7 years, 6 days ago)

Quote:

Orbit said:
Most likely the sryinge. But why not just put a drop on the plate and streak with a sterilized a loup. You can just use a piece of wire after flaming.



Yeah this would work as well and I will try it. Just with not the same syringe. Lol.


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OfflineDactylium
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Re: What a contaminated plate!! [Re: TheCyndicate]
    #24036899 - 01/24/17 06:31 PM (7 years, 6 days ago)

Why not try the syringe without the q tip just to be sure?


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OfflineOrbit
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Re: What a contaminated plate!! [Re: Dactylium]
    #24036922 - 01/24/17 06:40 PM (7 years, 6 days ago)

Might as well let the plates go for a while, some mic may pop up. Ive been battling bacteria laden spores as well.


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InvisiblebodhisattaMDiscordReddit
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Re: What a contaminated plate!! [Re: Orbit]
    #24036978 - 01/24/17 06:57 PM (7 years, 6 days ago)

Make an inoculation loop and make a streak plate like in my agar tek


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OfflineTheCyndicate
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Re: What a contaminated plate!! [Re: bodhisatta]
    #24037024 - 01/24/17 07:18 PM (7 years, 6 days ago)

Hey bod, I'm going too just figure sterile swab would work too. I have tossed all the contaminated plates. I could have saved them and cleaned them up but the bacteria showed up so damn fast and the red  plates in my other thread are doing fucking wonderful. And I just Resurrected my old ATL #7 culture and have that on agar as well . It has recovered nicely and I will make a grow log when it's ready.


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OfflineOrbit
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Re: What a contaminated plate!! [Re: TheCyndicate]
    #24037336 - 01/24/17 09:30 PM (7 years, 6 days ago)

I used Qtip storage for some spores. Infortunatly after 16 years they are titrs. I had no comtam or growth. Same for the plate. So its plausible that it may work.


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InvisibleMad Season
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Re: What a contaminated plate!! [Re: Orbit]
    #24037533 - 01/24/17 11:19 PM (7 years, 6 days ago)

Swabs can be PCd, no? Did OP PC them? If so it's gotta be the syringe, if not.. well..... I'd just get a loop regardless tho lol. Shit is the tits


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InvisiblebodhisattaMDiscordReddit
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Re: What a contaminated plate!! [Re: Mad Season]
    #24037619 - 01/25/17 12:26 AM (7 years, 6 days ago)

Quote:

bodhisatta said:
Quote:

bodhisatta said:
Id put some water in the jar with the swabs. I'd worry they're not sterilized since they're only in contact with hot air likely.

Water makes 1700 volumes of steam so it would only take a few drops in the jar to fill it with steam. During the cycle



wrong forum, lol fuck



https://www.shroomery.org/forums/showflat.php/Number/24037039#24037039


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OfflineOrbit
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Re: What a contaminated plate!! [Re: Mad Season]
    #24038077 - 01/25/17 08:58 AM (7 years, 5 days ago)

Quote:

Mad Season said:
Swabs can be PCd, no? Did OP PC them? If so it's gotta be the syringe, if not.. well..... I'd just get a loop regardless tho lol. Shit is the tits





He did PC them. They are probly pretty sterile out of a new box. You are working with a syringe which is non sterile to begin with. A loup would be the best choice though.


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InvisiblebodhisattaMDiscordReddit
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Re: What a contaminated plate!! [Re: Orbit]
    #24038078 - 01/25/17 08:59 AM (7 years, 5 days ago)

To sterilize swabs and other dry things they're usually put in an autoclave bag so steam can penetrate


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InvisiblemushboyMDiscord
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Re: What a contaminated plate!! [Re: bodhisatta]
    #24038180 - 01/25/17 09:45 AM (7 years, 5 days ago)

i just put them in a pint jar with the tips loosely wrapped in foil.

look up how q-tips are made. they are not sterile. shit they are sealed in thin cardboard.


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