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Ghost



Registered: 07/26/15
Posts: 25
Last seen: 2 years, 10 months
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Floating Pizza LC Jars & Testing without agar
#27129493 - 01/05/21 11:21 PM (3 years, 1 month ago) |
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So I am assuming that a floating pizza on top of my liquid culture is a good sign of contamination? I’ve gone years and years without messing with agar but I suppose the time has come. Just ordered some agar agar and 25 disposable Petri dishes and gonna have to learn how to use it. The way I’ve had to test this batch since I got a wall of lc’s going that date back at least a year is to whip up some PF style BRF cakes and start knocking them up systematically.... so I bought a labeled. I took 6 LC jars out to my Flowhood and labeled them A-F then covered it in clear tape so it didn’t rub off with alchohol. Then I filled the 6 syringes one per LC jar and put a letter on each syringe correlating to the jar it came from. Then I knocked up 6 BRF jars and did the same thing putting label A to the jar knocked up with syringe A and so on.... so now I know which jars of LC are contaminated. Three of them took off, looked and smelled fine and went into the Martha. The rest stalled or turned green.
This lovely green specimen came from this “floating pizza” jar and
This stalled out and 2 shades of brown beauty here came from this jar of milky ass clouds 
Yeah lessons learned... or learning anyway. I’m still wondering what r the benefits of adding an additional step (ms spore syringe -> LC -> grain master jar -> filter patch bag -> monotub vs. what ms spore syringe to ? Agar Petri dish? Just squirt it on the plate or how’s that work? -> LC or grain jar? -> filter patch bag -> monotub)
And can I just assume a floating pizza to always b contam and just throw it out without testing? And what’s the preferred process flow anyway? I’ve got new spore syringes on the way and gotta get this one right. Anywhoo.... I just felt a need to share my misery because it was getting lonely and wanting company I suppose
-------------------- Fine. One more one more time but then that's it!
 
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Josex
#cheat_code



Registered: 11/13/15
Posts: 8,995
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Re: Floating Pizza LC Jars & Testing without agar [Re: 1more]
#27129509 - 01/05/21 11:38 PM (3 years, 1 month ago) |
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That floating pizza stuff is what happens when you have a grown ass LC that's been sitting way too long on a shelf (or wherever you had it). It happens both to clean LC's and bad LC's. Your LC's just happened to have the floating pizzas and they also happened to be contaminated, but the two things don't correlate.
You can't inoculate an LC with spores and expect good results. You might get lucky but that's usually not the case because spores are inherently dirty and spore slution may contain mold spores and bacteria. An aqueous media is like heaven for bacteria.
Quote:
Yeah lessons learned... or learning anyway. I’m still wondering what r the benefits of adding an additional step (ms spore syringe -> LC -> grain master jar -> filter patch bag -> monotub vs. what ms spore syringe to ? Agar Petri dish? Just squirt it on the plate or how’s that work? -> LC or grain jar? -> filter patch bag -> monotub)
Yea too many steps. Put your spores on agar first and forget about liquids until you get your shit together. If you then wish to work with LC's the first link in my sig is all I do and it's very easy.
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