Atp

Time

Time

Apyrase

Apyrase dNDP + dNMP + phosphate ADP + AMP + phosphate

■ Figure 10-15 Pyrosequencing is analysis of pyrophosphate (PPi) released when a nucleotide base (dNTP) is incorporated into DNA (top left). The released PPi is a cofactor for ATP generation from adenosine 5' phosphosulfate (APS). Luciferase plus ATP converts luciferin to oxyluciferin with the production of light which is detected by a luminome-ter. The system is regenerated with apyrase, that degrades residual free dNTP and dATP (Step 3). As nucleotides are added to the system one at a time, the sequence is determined by which of the four nucleotides generates a light signal.

Nucleotide sequence

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