Product Number: T0956
Our Universal Transfection Reagent is a unique formulation of a proprietary polymer blend. Universal Transfection Reagent is used for transient and stable transfection of nucleic acids into various eukaryotic cell lines and hard-to-transfect primary cells. This fast and easy protocol is compatible with serum, serum-free medium, and antibiotics.
The Universal Transfection Reagent is provided as 1mL liquid in a single vial. This volume allows for either:
Universal Transfection Reagent can be stored at 2-8 °C for up to three months. For long-term, store at –20 °C. Allow all kit components to thaw and equilibrate to room temperature before use.
The procedure stated below is designed for the transfection of common cell lines with 1 µg/µl DNA plasmid (diluted in sterile molecular biology water). Culture and transfect cells in standard serum-containing or serum-free medium appropriate for the cell type. Use good aseptic technique and use only sterile materials.
DNA plasmids should be high-quality, ethanol-precipitated, resuspended in molecular biology grade water to a final concentration of 1 µg/µL. Optimal amount of Universal Transfection Reagent used depends on cell type and is generally 1 – 3 µL per ug of plasmid DNA.
Presence of serum (>5%) and antibiotics does not inhibit transfection and in some cases increases the efficiency of transfection. Transfecting cells in the presence of serum minimizes the toxicity.
This protocol can be optimized for use with a wide variety of cell types. Seeding density, amount of DNA used, and incubation time can easily be varied to achieve higher expression and lower toxicity when needed.
Day 1: Plate Cells
Plate the cells 18-24 hours before transfection. An appropriate seeding density should be used so the cell culture plate is 90-95% confluent at the time of transfection. Serum-containing or serum-free medium appropriate for the cell type can be used for culturing the cells.
Day 2: Transfection
Day 2: Optional Media Change (Sensitive Cells)
A complete media change can be performed 5 - 24 hours after transfection for very sensitive cells. For most cells, we recommend the media be changed only at 48 hours post-transfection until protocol optimization requires this extra media change.
Day 4: Change Medium and Check Transfection Efficiency
Day 1: Plate Cells
Plate the cells 18-24 hours before transfection. A seeding density of 0.5 - 1.0 x 106/mL should be used so the cell culture plate is ready at the time of transfection. Serum-containing or serum-free medium appropriate for the cell type can be used for culturing the cells.
Day 2: Transfection
Day 4: Change Medium and Check Transfection Efficiency
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