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Daddy4487
The creator

Registered: 08/18/20
Posts: 406
Last seen: 11 months, 1 day
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Help with LC?!
#27310514 - 05/15/21 10:46 PM (3 years, 7 months ago) |
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Hey everyone I had a quick question about LC I was hoping to get a little help with... this is my 3rd attemp making LC and the most recent attempt I even used my flowhood, I’m sure my culture is clean. And I made the LC using 500ml water and added 20 grams of 100% pure honey and mixed it well before pc ing at 15 psi for 13-14 minutes.... colonization is clearly forming but it seems to have trich or some kind of contam here’s some pics... any help would greatly appreciated
-------------------- Hope I can help
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Thanks for the help!
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Goatrider
Rhythm Guitarist



Registered: 04/08/20
Posts: 4,801
Loc: Germany
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You tested it on agar, got mold, and are sure your culture is clean?  Inoculated lc with clean agar myc? Pc vented properly? Are these possible inoc points or accidently hovered over the plates? Honestly if it doesn`t work for you i wouldn`t hassle around, and just make up some li. Maybe spend a pic of your lc if there`s some left.
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DERRAYLD
Constructus


Registered: 05/13/02
Posts: 12,109
Loc: South Africa
Last seen: 1 hour, 25 minutes
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That's definitely not a clean lc.
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Salvitica
Stranger


Registered: 04/27/21
Posts: 45
Last seen: 3 years, 3 months
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If a clean LC produces mold on agar, then something tells me the LC is probably not clean... That or your inoculation technique is not sterile enough. Contamination can always be invisible, but will surely show up, like in these plates here.
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Daddy4487
The creator

Registered: 08/18/20
Posts: 406
Last seen: 11 months, 1 day
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Okay I was pretty sure my cultures were clean, I will post a picture of the cultures I used and also pics of the lc’s themselves in a minute thanks guys
-------------------- Hope I can help
&
Thanks for the help!
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Daddy4487
The creator

Registered: 08/18/20
Posts: 406
Last seen: 11 months, 1 day
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-------------------- Hope I can help
&
Thanks for the help!
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Mateja



Registered: 07/14/16
Posts: 7,948
Loc: Here
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Quote:
Salvitica said: If a clean LC produces mold on agar, then something tells me the LC is probably not clean... That or your inoculation technique is not sterile enough. Contamination can always be invisible, but will surely show up, like in these plates here.
That statement is kind of the opposite of truth. Determining the presence of any kind of bacillus in liquid media is so straightforward and simple that you can show a 5 year old child how to do this with staggering precision. You can read for yourself and examine the photos I post in the LC Training Camp as I demonstrate over and over not just how predictable this examination method is but I try also to point out that dropping LC onto standard agar pucks should not be regarded as predictable at all since most bacteria will be greatly incapacitated or not be able to germinate when th aW is so low, hence why special 'motility agar' is made to test specifically for bacterial activity, besides this semi solid agar media a liquid broth is by far the most important growth media for propagating bacillus of any kind. My tip for inspecting a broth is first determine that bacillus isn't present (simply look at it for 3 sec, if broth turns turbid overnight then it's bacterial but if its as clear and transparent as its always been then you know it's not bacterial so now you only need to test for mold if something has occurred that would indicate possible late germination or other vector, but for the vast majority of time cube myc is pretty easily distinguished in shape, size and rate of growth from most contaminant molds.
-------------------- Cakes inside Water Tub
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Mateja



Registered: 07/14/16
Posts: 7,948
Loc: Here
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Quote:
Daddy4487 said:


I don't see turbidity just a clear broth with little to no light diffusion so it's guaranteed not bacterial and seems to contain 1 type of organism and not several different looking types so chances are pretty big that you got yourself a clean and fully colonized LC at this point. No stirring?
-------------------- Cakes inside Water Tub
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Daddy4487
The creator

Registered: 08/18/20
Posts: 406
Last seen: 11 months, 1 day
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Re: Help with LC?! [Re: Mateja]
#27314121 - 05/18/21 06:11 PM (3 years, 7 months ago) |
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Yea I stir it usually daily, actually have been contemplating getting a magnetic stirrer. But yea I didn’t want to stir it before I took the pic in case cloudiness may have altered anyone’s judgment/perception
Say hypothetically it’s a clean LC... what’s the best things I can use it for??
-------------------- Hope I can help
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Thanks for the help!
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