Droplet Digital PCR for AAV Quantitation
Type
Blog Post
...droplet in each well (Figure 2). A clean ddPCR should have a clear separation between positive (blue) and... to 2 minutes and the denaturation time to 1 minute has been shown to increase droplet separation (Witte... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...getting a clean separation between positive and negative droplets with these parameters, here are a few...Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the ... for ddPCR set-up.
Prepare the master mix in a separate area from where you prepare your sample dilutions...your PCR
For a good starting place on what PCR parameters to use, see Lock’s seminal paper on AAV titration...