A computer-readable medium containing instructions that, when executed by a processor, cause the processor to control an EWOD device to perform a method of performing nucleic acid amplification via polymerase chain reaction (PCR) using an electro-wetting on dielectric (EWOD) device, the PCR method comprising the steps of: receiving a liquid droplet within an EWOD channel defined by the EWOD device, the liquid droplet containing a nucleic acid; performing a PCR cycle comprising: generating a first thermal zone at a first spatial location within the EWOD channel, the first thermal zone being controlled to have a first temperature (T1) for performing a denaturing step, and moving the liquid droplet to the first thermal zone ; denaturing the nucleic acid in the first thermal zone ; generating a second thermal zone at a second spatial location within the EWOD channel different from the first spatial location, the second thermal zone being controlled to have a second temperature (T2) for performing a step of annealing primers, and moving the droplet to the second thermal zone; annealing primers of the nucleic acid in the second thermal zone ; while the liquid droplet is in the second thermal zone , re-equilibrating the temperature of the first thermal zone to a third temperature (T3) for performing nucleic acid polymerization, and moving the droplet to the first thermal zone ; and polymerizing the nucleic acid in the first thermal zone ; and performing multiple PCR cycles by repeating the PCR cycle until the nucleic acid amplification is complete. receiving a liquid droplet within an EWOD channel defined by the EWOD device, the liquid droplet containing a nucleic acid; performing a PCR cycle comprising: generating a first thermal zone at a first spatial location within the EWOD channel, the first thermal zone being controlled to have a first temperature (T1) for performing a denaturing step, and moving the liquid droplet to the first thermal zone ; denaturing the nucleic acid in the first thermal zone ; generating a second thermal zone at a second spatial location within the EWOD channel different from the first spatial location, the second thermal zone being controlled to have a second temperature (T2) for performing a step of annealing primers, and moving the droplet to the second thermal zone; annealing primers of the nucleic acid in the second thermal zone ; while the liquid droplet is in the second thermal zone , re-equilibrating the temperature of the first thermal zone to a third temperature (T3) for performing nucleic acid polymerization, and moving the droplet to the first thermal zone ; and polymerizing the nucleic acid in the first thermal zone ; and generating a first thermal zone at a first spatial location within the EWOD channel, the first thermal zone being controlled to have a first temperature (T1) for performing a denaturing step, and moving the liquid droplet to the first thermal zone ; denaturing the nucleic acid in the first thermal zone ; generating a second thermal zone at a second spatial location within the EWOD channel different from the first spatial location, the second thermal zone being controlled to have a second temperature (T2) for performing a step of annealing primers, and moving the droplet to the second thermal zone; annealing primers of the nucleic acid in the second thermal zone ; while the liquid droplet is in the second thermal zone , re-equilibrating the temperature of the first thermal zone to a third temperature (T3) for performing nucleic acid polymerization, and moving the droplet to the first thermal zone ; and polymerizing the nucleic acid in the first thermal zone ; and performing multiple PCR cycles by repeating the PCR cycle until the nucleic acid amplification is complete. <EOS>