Description of the Ovation Target Enrichment System.
(A) Experimental steps of the assay and time required for each step. Adaptors (green) are ligated on to generated double stranded cDNA (ds-cDNA). Probes (shown in red and yellow) are hybridized to target cDNA and extended with a polymerase (dashed grey lines). All probes have common tail sequences (blue), which are used as priming sites along with adaptor sequences in subsequent library amplification PCR steps. (B) Example of probe positioning across different exons in a full length double stranded cDNA. Each exon (demarked by blue vertical lines) will have probes (green arrows; arrow points in the 3’ direction) designed to hybridize near the predicted exon-exon junctions. Exons larger than 300 nucleotides (nt) may have additional probes tiled along the length of the exon to obtain more complete sequence coverage. Probes are designed against both strands of the cDNA to enable identification of gene fusions when only one of the pair of genes is targeted. Translation start sequence (ATG) and poly A tail are labeled.