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Help: H4 Stable cell line using pLVTHM vector

H4 cells stable cell pLVTHM vector Flow cytometry

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#1 Carlo Leicester

Carlo Leicester

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Posted 10 January 2014 - 04:18 AM

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







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