Always use 25uL of input volume for your Crystal Digital PCR experiment. When you load the chips, make sure you do not carry any air bubbles in the pipet tip. When you prepare the PCR mixes, we recommend that you wait 2/3 minutes after the final vortex/centrifugation and before filling the chips’ inlet ports. As well, once the chips are loaded, wait 2/3 minutes before sealing the ports with the long white caps. This will allow potential micro-air bubbles to migrate up to the surface. Finally, if you observe air bubbles at the surface of the mix (before sealing), try to pop them with a pipet tip.