Fig. 3

Amplicon-based and hybrid capture sequencing methods. The figure shows a hypothetical gene for which a clinical assay sequences exons 2 and 3. The DNA is sheared either in recovery from being formalin-fixed and paraffin-embedded, or deliberately to allow for sequencing adapter binding. Hybrid capture involves probes that are designed with homology to the gene of interest and bind cDNA. Notice that the fragmented DNA can contain information beyond the boundary of the exon. The probes are biotinylated and unbound DNA is washed away. In amplicon-based sequencing, only probes that contain the complementary sequence for both primers are amplified. Therefore, no information outside of the primers is sequenced