high efficiency for B cell transfection

William F. Wade William.F.Wade at dartmouth.edu
Wed Aug 23 09:28:36 EST 1995


My lab is starting a project to transfect cDNA into a murine B cell
line (M12.C3) in the hope of changing the class II signaling phenotype
from PKC/cAMP to the Ca2+/tyr kinase phenotype associated with primed
muring B cells.  We typically transfect M12 cells by electroporation
and get 1 in 10,000 cells transfectanted, and 80% express the genes of
interest.  Even transfecting 10 million cells the number of cDNA we
could analyze will not be great using them method.

The question, does anyone have, or know of a method for transfecting
murine B cells that has a higher efficiency?  I have heard there are
methods that approach 100%.  Would spheroplast fusion, ala, Brian
Seed's expression cloning method get there?  Will it work with
lymphocytes?

Any advice or ideas are appreciated,my email address is
william.wade at dartmouth.edu

Thanks for the help
Sincerely,
William F. Wade, Ph.D.
Assistant Professor
Dartmouth Medcial School



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