*2.6. Complexation of the rAAV Vectors with the Carbon Dots and Release Studies*

The various CDs (40 <sup>μ</sup>L) were directly mixed with the rAAV vectors (40 <sup>μ</sup>L, 8 <sup>×</sup> 105 transgene copies) and incubated for 30 min at room temperature to generate the rAAV/CD systems. Alternatively, Cy3-labeled rAAV vectors were employed for the visualization analyses of the complexation studies by mixing Cy3-labeled rAAV (40 <sup>μ</sup>L, 8 <sup>×</sup> 10<sup>5</sup> transgene copies) with the CDs (40 <sup>μ</sup>L) in 96-well plates in serum-free DMEM (100 μL). Cy3 labeling of the samples was monitored under live fluorescence with a rhodamine filter set (Olympus CKX41, Hamburg, Germany). For the release studies, the rAAV/CD systems as prepared above were placed in 24-well plates in 350 μL of serum-free DMEM, and rAAV was measured in aliquots of culture medium at the denoted time points using an AAV titration ELISA [25].
