Transfection by electroporation of 293 suspension cells - electroporation of 293 cells for large scale transient expression (Oct/24/2006 )
Hello, I have been trying to electroporate 293 cells for large scale transient expression. I am using Freestyle from Invitrogen as my culture medium and Earl's buffer as the salt neutralising buffer. I am applying a voltage of about 160-190V to 800ul of cells/DNA mix. The cells are initially seeded at 1-2x10^6 cells/ml. However, after electroporation the cells are dying more quickly than expected. Does anyone has any experiance with electroporationg 293 cells in suspension. Thank You for the help.
I think linear PEI is a good alternative to transfect large scale 293 cells.
Hello
Thanx for your response. I do use PEI as well but certain proteins express much better by electroporation that's why am trying to electroporate 293 cells. I have achieved a fair amount of success but I am looking for ways to maximise my protein production.
Hello, I have been trying to electroporate 293 cells for large scale transient expression. I am using Freestyle from Invitrogen as my culture medium and Earl's buffer as the salt neutralising buffer. I am applying a voltage of about 160-190V to 800ul of cells/DNA mix. The cells are initially seeded at 1-2x10^6 cells/ml. However, after electroporation the cells are dying more quickly than expected. Does anyone has any experiance with electroporationg 293 cells in suspension. Thank You for the help.
I think linear PEI is a good alternative to transfect large scale 293 cells.