Viral Gene Delivery and Evaluation by Flow Cytometry

This 12-page resource, from Frederick Community College, is a standard operating procedure (SOP) for viral gene delivery and evaluation by flow cytometry. The SOP outlines the purpose, scope, definitions, references, safety measures, responsibilities, materials, and equipment involved in the procedure. Step-by-step procedures are included for transduction of mammalian cells with the gene for green fluorescence protein (GFP) using adeno-associated virus (AAV) vector and subsequent evaluation of transduction efficiency using flow cytometry.

Procedure steps include: transduction preparation, transduction of HEK293 cells, cytometer setup and calibration, starting up the cytometer, preparing for instrument QC, running instrument QC, preparation of transduced samples, setting up the acquisition parameters on the cytometer, creating sample gating strategy on the cytometer, sample acquisition, data analysis, and exporting the data. Detailed steps are included.