Hi Guys, I am pretty new to here and also in using cells. In part of my project I have to generate some stable cell line using lentiviruses. I cloned and genereate these lentiviruses using pLVTHM vector. Indeed I need to downregulate some genes of interest. The pLVTHM vector does not have any antibiotic resistance thus I can use this strategy for selecting cells and generate stable cell line.
So the question is: How can I do?
Tecnically, I understood that I have to infect the cells and then sort them using Flow cytometry.... what next? Will be sure that the cell sorted will be stable forever or do I need to sort them after a week ect?
Does anyone have a protocol to follow?
Thank you very much guys any help will be super appreciated