Quantification of Molecular Transfection Efficiency

GFP is often used as a control to determine transfection efficiency of a transfected cell line with a particular gene of interest, this efficiency is easily calculated using FLOW.

  1. After transient transfection and incubation, acquire 1x105 and 1x107 cells from incubator and place into a 15mL conical tube. Spin at 1200rpm for 5minutes (detach adherent cells using Trypsin if needed).
  2. Aspirate supernatant and resuspend cells in 300-500µl of cold FACS buffer.
  3. Send samples to AVIVA for flow cytometry analysis *Commonly used F ACS buffer recipe: 1X PBS; 2-10% FBS; 0.5-1% BSA; 1mM EDT A; *0.05% Sodium Azide. (*Warning: Sodium Azide will inhibit metabolic activity)