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Next Subject 		Using LB Broth to check for contaminationemma542008-04-30Click here to register.
Hi, I've been having problems in my lab with contamination and I have been told by my supervisor to check my media, trypsin etc. for contamination by putting them in a universal with LB broth and shaking them. She gave no other guidelines and now I cant get hold of anyone to help me. I was wondering how much LB broth to use and how much media, and also how long to I need to shake the mixture for?

Thanks
 
Re: Using LB Broth to check for contamination
sferg2008-05-05Click here to register.
to check for contam, we put a TC dish containing our culture media alone in TC incubator (i.e. 7-10ml into a 100mm TC dish). you can have additional plates containing media that has been inoculated with a small amt (50-100ul) of your trypsin, PBS, ect to the media to test for your source of contam.
 
Re: Using LB Broth to check for contamination
n@@n!2008-05-06Click here to register.
To check media and other liquid components used in cell culture, better to follow the standard protocol, what "ferguson" has suggested. Besides there are other sources of contamination like the laminar flow - check the condtiion of the filter - you can do this by simply keeping few petridishes with microbial growth media in opened condition and run the flow for few minutes, and then take out, incubate in conventional incubator. One more thing is Co2 incubator - better you disinfect the system by following standard protocols, like fumigation (in worst cases) or simply wiping with Ethanol etc.

But in cell culture its better to keep in mind that PREVENTION IS BETTER THAN CURE.

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