Thp1 transfection

Guriq Basi gbasi at SLIP.NET
Wed Feb 19 19:44:02 EST 1997


help!  i am trying to obtain stably transfected Thp1 (Thp-1/Thp 1)
myelomonocytic cells (human), without success.  has anyone out there
ever done this and succeeded?  please reply with your detailed protocol.

i am using electorporation (10-20 µg DNA), with a biorad gene pulser at
220V/960 µF, and within 48-72 hrs post transfection, *98% of the culture
is dead.  add drug (G418) and wait 2 wks., and nothing grows out.  i am
pretty convinced (from control expts.) that the transfection alone is
killing the cells.  transients with ß-gal reporter indicate decent
transfection efficiecy (ca. * 10%) under these conditions.  thank for
your help.

gbasi at slip.net



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