I use electroporation method for transfection. I want to know the factors on which the transfection efficiency depends and what measures can be taken to improve the efficiency?
Some excellent commentary is available here:
http://www.bio-rad.com/webroot/web/pdf/lsr/literature/10-0826_transfection_tutorial_interactive.pdf
I use electroporation for mammalian cell transfection. But it causes a lot of cell death. I start with high number of cells but it has not helped much. Can anyone please provide any solution to this problem? Also I need suggestions to improve the transfection efficiency. Thanks!
Which cells are you transfecting? Certain cells (lines) can be easily transfected using chemical based methods such as calcium phosphate, PEI etc. Electroporation is usually hard on cells but it can be fine tuned and an acceptable compromise between cell death and transfection efficiency can be achieved. Other alternatives include the use of Amaxa's Nucleofector and Invitrogen's Neon system. I have used both of these systems to transfect hard-to-transfect T cell lines and primary cell lineages to very high efficiency (up to 80%). You need to give more details.
I use U2OS cells, and I have no other option than electroporation. I have to transfect the cells twice within 24 hours. After first transfection I allow cells to adhere for 12 hours and then change media, then after another 12 hours I do the second transfection. So, cell death is quite inevitable. But I wanted to minimize it.
- Badges
- Science method