Cell Preparation for Transfection
Plate cells approximately 24 hours before transfection, making sure cells are at optimal concentration (70–90 % confluency).
Steps Before Transfection
- Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti:MEM®I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
- Place diluent in a sterile tube.
- Add plasmid DNA. Gently pipette up and down to mix.
- Add X-tremeGENE™ HP DNA Transfection Reagent to the diluted DNA.
- Incubate for 15 minutes at +15 °C to +25 °C.
- Add transfection complex to the cells in a dropwise manner.
- Gently shake or swirl the wells or flasks to ensure even distribution over the entire plate.
- Incubate cells for 18 – 72 hours before measuring protein expression.