293 Cells: how to transfect?

WB brune at sun0.urz.uni-heidelberg.de
Mon Jun 24 22:28:00 EST 1996


jstrassw at opal.tufts.edu wrote:
> 
> Hello!
> I am trying to use 293 cells because they are reported to be highly
> transfectable (>50% by Calcium Phosphate) and they are EGF-responsive.  The
> only problem is that the buggers dont adhere well to dishes.  I am loosing a
> lot of cells every time I wash after transfection.  Any suggestions?
> 
> Thank you,
> John

John: You are right, 293 cells don't adhere firmly to culture dishes. To
avoid losing cells, try to pipet fluids (medium or PBS) CAREFULLY / 
SLOWLY
into the edge of your culture dish. If you pipet fluids forcefully onto
your 293 cells they will get washed away, especially if cells are 
confluent.
(BTW, I had the same problem when I started working with these cells.)
Good luck!
Wolfram



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