2.3.2. Quantitative siRNA Encapsulation Efficiency (EE%)

The siRNA encapsulation and retention of all liposome preparations was determined using the Quant-iT RiboGreen RNA Assay Kit (Thermofisher Scientific). Following dialysis purification from un-encapsulated and/or un-retained siRNA, liposomes were denatured and solubilized in 1% Triton X-100 at 37 °C for 15 min to release siRNA cargo. Released siRNA was mixed with RiboGreen reagent for fluorescent labeling and emission was read at 525 nm. Fluorescence units of solubilized liposomes was fit to a known standard curve of siRNA in 1% Triton X-100. EE% of each liposome formulation was calculated as (pmols siRNA encapsulate/total original pmols siRNA used) × 100.