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TransPass R1: Plasmid DNA and siRNA Transfection Protocol |  | | TransPass R1: Plasmid DNA and siRNA Transfection Protocol |
Protocol
- Plate cells at an approximate density so that they reach 60-70% confluence the day of transfection.
- Mix an appropriate amount of serum-free medium (Table II) with plasmid DNA in a sterile tube and mix by vortexing. Add an appropriate amount of TransPass D1 DNA Transfection Reagent for the size of plate you are using (Table II). Incubate at room temperature for 20 minutes.
- Aspirate culture medium from cells. Wash cells once with serum-free medium. Aspirate the culture medium from the cells and immediately replace with the transfection complex mixture.
- Incubate cells for 3 hours.
- Replace medium with fresh medium containing serum. Incubate cells 2-3 hours.
- During this time prepare the siRNA transfection complex according to protocol 1.
- Aspirate the culture medium from the cells and immediately replace with the diluted siRNA transfection complex mixture (step 4, protocol 1).
Incubate cells 24-48 hours before analysis.
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