I'm working on HuCCT-1 a cholangiocarcinoma cell line. I'm trying to transfect cells with shRNA for CXCR7 but I did not have succes with lipofectamine. Can you give any suggestion for any successful technique? Thanks a lot
Thanksfor your answer. The problem is that shRNA of GFP did not enter inside my cells with lipofectamine method. We also tried with lentiviral transduction without any great results.
Hi Alessandra,
to be success in transfection of shRNA in to a cell some steps you should consider. First, The confluency of prepared cells( i preferrred 50-60%). second, control group. you could do this by transfecting GFP alone ( not shRNA). it will show you whether using the same protocol is able to transfect the cells or not. If you don't get transfected cells ( just view them under microscope) with the GFP, you should check your status of your reagent. may be need to change it.
Good Luck,
you could try using Fugene6 transfection reagent which at times works pretty good in difficult to transfect cells
Thanks a lot to everybody for your contribution.I finally got transfection with shRNA for RDF using the program T020 of amaxa. I'll try in the next few days to try this tecnique for CXCR7 shRNA . Is there someone here that can tell me for how long plasmids remain inside the cells silencing the gene that I'm interested in? Thanks again
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