In brief i collect the tissue from research lab and by follicular puncture method I collect the fluid.By centrifugation I collect the cells and subjected to ficoll gradient centrifugation. I culture the intreface in DMEM medium with 10%FBS and 1% antibiotics. In microscopic examination i came to know that media is heavly contaminated with bacillus and cocci bacteria. Kindly help me in this
I had this problem in two different labs where I had a privilege to have my post-doc training. After many different experimental variations that we tried to test we found that the contamination most likely occurs at the step of removing the mouse ovaries from the animals (it was coming from skin hair of mice). The solution was very simple, before any surgical invasion (after you euthanized the mouse) you need to completely soak the whole mouse body in 70% Ethanol for 3 min. After that you can start your surgical procedures. It sounds like a very primitive solution but, believe me, the result was very efficient.
Thank you sir .
I will include this step in my protocol.sir do you use any antibiotics in media apart from normal antibiotics for primary cell culture and any other ingredients in addition to FBS.
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