FIG. 2.

Mutation detection using the F1 Pyrosequencing primer. (A) Wild type; (B) CTG₅₁₁CCG substitution. The Pyrosequencing chemistry differs from Sanger sequencing. In the former method, each nucleotide is dispensed automatically one at a time. Whenever a nucleotide is incorporated, a flash is produced and is monitored as a peak in the pyrogram. The height of the peak is proportional to the number of nucleotides inserted. Mutations are detected both at the peak level and at the aligned sequence output.