For difficult to transduce adherent cell lines, I was wondering if anybody has tried spinoculation with polybrene? If so, what is the protocol you used?
I usually used 8-10 ug/ml polybrene in growth media, using 1ml per well in 6 well plates. Some protocols I went as high as 16 ug/ml. Check literature for info about your particular line, or titrate.
I’d infect late afternoon and change media to growth media the next am. I’d usually assess success at 72 hr post infection, in my case by GFP positive cells. Hope this helps and best
of luck!
Susan C Hubchak Hi Susan, thank you for your response. Have you tried spinning any adherent cells with polybrene before? If so, what was the recommended time of spinning and conditions?
I have not tried that as we did not have the proper centrifuge attachment.
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