We work with WIL2 cell line and want to trasfect mirna mimic and inhibitor. We tried first with lipofectamin and than electroporation (using amaxa nucleofector, U-001 program), none of that works. medium culture: RPMI 10% FBS.
Hi Serena, I've been working suspension cells for years. For sure, lipo-transfection never works in suspension cells. I've tried electro-transfection before, it works but has very low transfection efficiency (around 20-30%). The most efficient way to introduce plasmid is to use viral-mediated transfection (lenti- or retro-virus).
Hope this information is helpful for you.
Hey,
As Han already said, I generally wouldn't recommend transfection, use lenti if possible (easier to work with over retro). It's easy to make your own as long as your lab has clearance and I can send you my current protocol if you want.
You can try using PEI. It is cheap and efficient. Here is a ref protocol that I followed for my suspension HEK cells (http://cshprotocols.cshlp.org/content/2008/3/pdb.prot4977.full) but this might work for your cells as well.
Hallo Serena,
I would recommend LipofectamineRNAiMAX. If you are performing a forward transfection, Please try reverse transfection and If possible optimize your transfections with Opti-MEM® I Reduced Serum Medium.
https://www.thermofisher.com/de/de/home/references/protocols/cell-culture/transfection-protocol/rnaimax-reverse-transfections-lipofectamine.html
BestHarsha
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