An investigation was conducted to determine the suitability of electrowetting-based droplet actuation for microfluidic PCR applications. We experimentally verified that droplets containing standard PCR reagents could be microactuated with the same facility previously demonstrated for droplets of deionized water or salt solution. We further verified that the manipulations and the environment inside the electrowetting chip did not inhibit PCR and we were able to successfully perform real-time PCR assays in 300 nL droplets within the electrowetting chip.