Figure 1.

Schematic representation of targeted next-generation sequencing and annotation for tumor specimens. Doubl-stranded DNA (dsDNA) is extracted from FFPE specimens containing tumor cells. Targeted regions are enriched by PCR and clonally amplified by emulsion PCR (emPCR). Such DNA fragments are sequenced by an NGS instrument and aligned to a reference genome. Variations from the reference are called by variant caller algorithms generating a VCF file. Rich variant annotation for somatic mutations can be achieved by querying multiple publicly available databases to create a meaningful clinical report for the management of oncology patients.