AAV titration with qPCR

Titration of AAV genomes in a purified sample is critical for ensuring accurate dosing. Titering AAV samples is typically accomplished by first treating samples with a DNase to degrade DNA not contained within the AAV capsid (i.e. unencapsidated DNA). Subsequently, the DNase is inactivated and encapsidated genomes are solubilized by denaturing and digesting the capsid. The number of DNase-resistant genomes in the treated sample(s) can then be absolutely determined using qPCR, with comparison to a standard of known concentrations. Alternatively, a droplet PCR method can be used, which bypasses the need for a standard. Here, we describe how to titer AAV samples using qPCR.