Impact of template addition volume and analyzed volume on digital PCR sensitivity

Digital PCR is a superior method to qPCR for the detection and absolute quantification of low concentration target templates. There are multiple digital PCR systems on the market that differ in numerous aspects including the amount of dead volume, which is the volume that is loaded but not analyzed by the given instrument. While it has been speculated that dead volume could impact the sensitivity of dPCR applications, here we provide data to support the conclusion that the most important factors in determining the relative sensitivity of each system are template addition volume and template analyzed volume. In summary, data provided herein demonstrate that higher template addition volumes can overcome any limitations that dead volume may have on the sensitivity of a dPCR application.