Hi
I'm trying to transfect my HeLa cells with addgene pSpCas9(BB)-2A-GFP (PX458) plasmid by CaP transfection method. However, after a long time of trial and errors, my transfection efficiency is very disappointing (I can see almost NO GFP).
We have controled everything; buffers, plasmids, cells, etc. everything is ok, but the results. Hope someone could help me with this problem.
I had the same problem transfecting them, but I tried different methods and achieved the highest efficiency by using electroporation.
Hope it helps you
Best wishes :)
Thank you Levin Joe Klages
I tried to optimize Electroporation for several weeks, however either it killed all my cells or didn't succeed in transfection! (Anything at all).
Anyway I solved it by using a commercial transfection reagent named as Transfectimine (http://darabiotech.ir/transfectimine/).
Thank you David Lee
for your advice. As you mentioned CaP is not a promising method and failed to transfect my HeLa cells. Plus, my plasmid size was relatively large (about 10 kb). However purchasing the product you suggested was not possible for me at the time (due to sanctions sanctions imposed against iran).Finally I used a locally available commercial transfection reagent (Transfectimine) and it solved my problem more or less.
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