Hello,
I am currently working on a project that requires me to transfect a DNA plasmid (5,600bp) into HVMECs. I want to inquire about available transfection protocols that would yield in high efficiency of transfected cells. I previously tried using chemical transfection reagent, JetPEI but results were very variable. My DNA plasmid has no reporter gene to measure transfection efficiency, and encodes a lncRNA so western blot analysis is not feasible. I am attempting to measure efficiency by looking assessing downstream genes via qRT-PCR. Any suggestions on better transfection protocols would be highly appreciated. Thanks in advance!
Hi, transfection can be done by electroporation and maybe ot can be more useful than chemical agents because of cell membrane potential.
Best
Hi, maybe you could just insert a reporter gene or selection marker into the expression plasmid of lincRNA you are interested in.
Or you may be interested in utilizing electroporation methods to transfect plasmid DNA, that could achieve relative high transfection efficiency. For example:
https://knowledge.lonza.com/cell?id=218&search=HMVEC
Or you may want to try lentiviral or AAV, that is viral vectors, to achieve stable and high transfection efficiency.
- Badges
- Science method