Notice how emulsion PCR is different from Illumina's bridge amplification, but serves the same purpose of copying the DNA.

This approach is similar to Illumina's reversible termination. However, parellelized pyrosequencing uses emulsion PCR, in which an environment of small water droplets is created inside an oil solution, instead of a solid surface as in reversible termination. Each "bead", or water droplet, contains a DNA fragment, and PCR of the DNA fragment takes place in each droplet. Then, similar methods of optical detection are used to sequence the DNA in each bead. Each modified base, when incorporated, releases a molecule called a pyrophosphate, which flashes detectable light in a series of enzyme-catalyzed reactions. (38) (48)

1. Incorporation of a modified base results in release of a detectable pyrophosphate (red box).

2. Pyrophosphate enters a reaction with AMP, resulting in an energy-rich molecule called ATP which reacts to release light. This burst of light can be detected and recorded for the base which is added.

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