I am working on mouse primary hepatocytes, and I need to quickly knock down some genes, to allow for evaluation of their function in xenobiotic metabolism. Which transfection reagent gives high transfection efficiency? What are the best media to use for transfection?
Yes, Mike
Lipofectamine RNAiMax is optimally transfect siRNA to several primary cultures. Do you really think that the antibiotic (pen/strep) has any impact on transfection frequency? I hardly find any thing.
Mike: I have seen similar procedures with most of the transfection agents but I really do not find any difference. Between, have you tried with Lipofectamin 3000? My transfection efficiency looks better with this reagent.
Thanks a bunch, Mick Frissen and Venkatesh Rengaswamy. The standard procedure is to culture the cells overnight (after isolation), can there be any effect on cell viability if I try the KD earlier, say 4 h post plating?
Hi Hebson,
you should also consider another technology instead of Lipofection that is called Magnetofection. This magnetic transfection method has been very successful in a lot of primary cells, here is a review that highlights this method
http://www.ncbi.nlm.nih.gov/pubmed/21893135
you can also find information here: http://www.ozbiosciences.com/content/13-magnetofection including citations database and cell database.
Good luck
Hi,
for siRNA transfection of primary hepatocytes I use Viromer BLUE (from Lipocalyx). I get up to 100% eficiency: https://www.ssl-id.de/www.lipocalyx.de/applications/rnai
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