Are you new to transfection and don’t know where to get started? Our team of Ph.D. educated Technical Support
Scientists have put together their top suggestions for transfection success. By following this workflow, you can
select the right reagents for your needs, then optimize conditions to achieve the best possible results.
Every cell line is different and may require specific transfection conditions. Therefore, it is important to
consider the following factors:
If cells are difficult to transfect with a lipid-based reagent and cannot be electroporated, there are other
options. Our Accell siRNAs are chemically modified to self-deliver into hard
to transfect cell types without the need for lipid-based transfection reagents or electroporation. Lentiviral
delivery of molecules is also a possibility for some applications.
Once you have selected the ideal delivery reagent for your cell type, it’s time to determine the optimal delivery
conditions. The primary determinants of a successful transfection are transfection reagent volume and cell density.
Test a range of lipid volumes using three different cell densities, using an appropriate positive control and
quantitative downstream assay, similar to seen in this Tech Note. Our product-specific delivery protocols provide guidance for initial volumes
of lipid to test. It is important to monitor cell viability using a resazurin assay (or similar), as lipid-based
transfection reagents can be toxic to some cell types. For most applications, an optimized transfection is
considered to be >80% delivery efficiency and cell viability.
By following these two steps, you will be able to pick the optimal transfection reagents for your studies and
determine the best conditions for your experiments. If you run into trouble or have any questions, please contact
our Technical Support team!
Authors: Ivonne Rubio Ph.D. and Kurt Marshall Ph.D. | Technical Support