DNA/Oligos delivery into the tough guys - RBL2H3's

Michael J. Tuvin mtuvin at mbcr.bcm.tmc.edu
Thu Mar 17 18:40:11 EST 1994


Hello, Netters!

I'm trying to deliver antisense oligos into RBL(rat basophilic leukemia)-2H3 cells
Those little .... don't seem to uptake anything alien from the bath, are not
affected by lipofectin (people say, haven't tried it myself) and aren't
responsive to the elecroporation. I use BioRad machine for the latter and tried
960uF in the range of 50 to 450 volts. Cuvettes 0.8 ml, width 0.4 cm.
Electroporation is conducted in RPMI media +10% FBS. Efficiency assessed by stayning with Trypan
(checking the dead/live ratio) and uptake of the fluorescent probe from the bath.
 No difference from the control.
 
Does anyone have any suggestions on the elecroporation conditions or alternative
method of delivery? 

If it isn't that big of a bother, please respond via e-mail to :
mtuvin at bcm.tmc.edu
                           Thanks.       MT



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