DNA-seq of human: adult male lung
DNA (0.1 mg) was extracted from the tumor tissues, and the formalin-fixed and paraffin-embedded tissues, followed by fragmentation. For the library construction, DNA (200 ng) was end-repaired and ligated with barcoded molecular adaptors, followed by PCR amplification to add indexes. ALK fusion was analyzed and annotated using the Omigen 101 genes pane. An in-house designed panel targeting most exons and selected introns in 110 cancer-related genes was used to capture DNA fragments to detect ALK fusions. In particular, hybrid capture-probes tilling the intronic regions of EML4 (intron 6, 13, 20), ALK (intron 1819), and KIF5B (intron 1516, 24) were designed for the detection of ALK rearrangements event. Sequencing libraries were generated after PCR amplification and then sequenced on DNBSEQ-T7 platform (MGI, Shenzhen, China) with 100 nt paired-end strategy.