VirusGEN AAV Transfection Kit – 1 Kit for 1 L of cell culture

Supporting Data

TransIT-VirusGEN® Outperforms Competitor Reagents in Suspension & Adherent AAV Cell Cultures. Suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium (A) or adherent 293T/17 cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio, 1.5 µg/ml, Agilent Technologies) with the following reagents: TransIT-VirusGEN® (2:1, vol:wt), Competitor B4 (3:1), Competitor B5 (3:3:1) or 25 kDa PEI (4:1, PolySciences). Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction using guava easyCyte™ 5HT Flow Cytometer. Functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the standard deviation of triplicate wells.

TransIT-VirusGEN® Enables Broad Scalability (AAV). Suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium (A) or adherent 293T/17 cells (B) were transfected with pAAV-hrGFP, pAAV-RC, and pAAV-Helper (1:1:1 ratio, 1.5 total µg/ml, Agilent Technologies) using the TransIT-VirusGEN® Transfection Reagent (2:1 reagent-to-DNA ratio (vol:wt), Mirus) at the indicated volumes perculture vessel. Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction by flow cytometry. Functional titers were measured from virus dilutions with less than 20% GFP positive cells.  Error bars represent the standard deviation of triplicate wells, flasks, or dishes.