Hi Guys,
I am trying to make a stable cell line from NIH/3T3 by transfecting with my plasmid of interest. After transfection, while I was changing the medium, I applied a bit harsh pressure on cell plate due to which a part of cell layer detached and now this layer is settled on top of monolayer in plate. I am suppose to add antibiotics to the plate now. Shall I dissociate the cells in single cells again (by trypsinization) and than add antibiotic or I can just add the antibiotic to plate and hope cells from both layers will die?
Please let me know at the earliest!
Best,
Ankur
its better trypsinize it and do it again. Since you are going to do fluorescent microscopy imaging, the monolayer will give better images
I would trypsinize the cells before adding the antibiotic to maximize exposure to the antibiotic. It won't add much time in the long run.
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