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Reverse transfection of siRNA

WS Won-Min Song
PA Praveen Agrawal
RI Richard Von Itter
BF Barbara Fontanals-Cirera
MW Minghui Wang
XZ Xianxiao Zhou
LM Lara K. Mahal
EH Eva Hernando
BZ Bin Zhang
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Transfection conditions were optimized for SKmel147 and A375 melanoma cell lines using fluorescein-labeled oligos (Block-IT, Invitrogen). Liposomal transfection complexes with siRNA pools for each of the candidate genes and with NTC (siRNA ON-target plus SMARTpools, Dharmacon, 50 nM) were generated with Lipofectamine 2000 (Invitrogen), following the manufacturer’s recommendations. Media was changed after 6 hr incubation with liposomal complexes. Forty-eight hours after initiation of transfection, cells were used for RNA extraction, or for proliferation or invasion assays.

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