HeLa S3 cells were transfected with a range of concentrations of siRNA targeted against CDC2 using HiPerFect Transfection Reagent from QIAGEN or Reagent L from another supplier. Non-silencing siRNA targeting green fluorescent protein (GFP) was also transfected. After 48 hours, CDC2 knockdown was assessed using quantitative, real-time RT-PCR. CDC2 expression was normalized to the expression of GAPDH. Expression levels relative to untransfected control cells are shown.
In reverse transfection protocols, cells are seeded and transfected in the same day. siRNA/miRNA is spotted into wells followed by the addition of HiPerFect Reagent. After complex formation, cells are added to the wells. These transfection protocols are rapid and convenient, can easily be automated, and are frequently used for high-throughput experiments.
Normal human embryonal keratinocytes were transfected with siRNA targeting lamin A/C or with AllStars Negative Control siRNA. Lamin A/C expression was measured by real-time RT-PCR after 48 hours.