Figure 1.

Schematic presentation of the principle of multiplex Pyrosequencing on a single template (left) and on multiple templates (right). PCR was performed with one of the primers biotinylated. Subsequent to immobilization of the product on streptavidin-coated magnetic beads and single-stranded elution, several sequencing primers were hybridized to the template(s) and Pyrosequencing was performed. A unique pattern was obtained, determining the exact sequence of different nucleotide positions.