I want to evaluate efflux pump activity using the Ethidium bromide-agar cartwheel method.
But in this method, I will have to prepare agar plates containing Ethidium bromide and inoculate bacteria. Question is, can I stterilize the plate afterwards?
Everywhere I am seeing that I should not autoclave anything with Ethidium bromide in.
Dear Shadid Uz Zaman
I will also suggest you the same, never autoclave carcinogenic reagents, it might produce harmful vapours. hence sterilize the agar (by autoclave), then by using syringe membrane filter sterilize the
Ethidium bromide (proper precaution is required). once the agar reaches the hand bearable temperature add the Ethidium bromide aseptically to the agar.
Dear Natarajan,
Yes, I have already prepared manual to prepare the agar plates in the way you suggested. That's not the main problem.
The problem is, once I have finished the test, the plate will have powerful MDR bacteria in it. But can I sterilize the plate before disposing it to biohazard bags?
Ethidium bromide destroy by autoclave , but you can sterilized their solution by millipore and added to agar before pouring
Ethidium bromide is carcinogenic, so it is preferable to dispose of the dish rather than sterilize it.
