Accurate determination of AAV genome copy titer is essential for reproducible dosing. Droplet digital PCR (ddPCR) provides absolute quantification without the need for standard curves, making it a preferred method for titering. This protocol covers the entire workflow from sample preparation, including DNase treatment and DNA extraction, to droplet generation, PCR cycling, and data analysis. We provide guidance on selecting appropriate primers and probes, as well as gating strategies to distinguish positive droplets. With careful attention to controls and replicates, ddPCR yields precise and reproducible titers for your AAV preps.