293 Cells: how to transfect?

Andy amcshea at world.std.com
Sun Jun 23 14:42:44 EST 1996


iddavis at vms.cis.pitt.edu wrote:
: In article <1996Jun21.142754 at opal.tufts.edu>, jstrassw at opal.tufts.edu writes:
: > 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?

Stick them down on collagen/poly lysine, dont leave them long at room 
temp, try and avoid washing them in PBS with not a lot else in it. I use 
lipofectamine at 5 microlitres to 1 microgram DNA and tranfect in the 
presence of 0.5% serum in DMEM overnight and get good efficiency (70%+). 
They are so transfectable its worth the lack of stickiness. Alternatively 
select for a sticker sub-clone of 293's (depends what you are trying to do).
Good luck,


			ANdy




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