Target-Capture of HBV integration in Hepatocellular Carcinoma
Genomic DNA was fragmented by sonication to yield 1001000 bp fragments, which were A-tailed and ligated to asymmetric DNA linkers.  HBV integration sites were then isolated by semi-nested ligation-mediated PCR using either forward or reverse primers specific to different HBV genomic region. Targeted sequences were further enriched by performing nested PCR with forward or reverse MiSeq HBV primers and forward or reverse MiSeq pLinkers, all containing an Illumina adapter for flow cell surface annealing. Forward and reverse libraries for the same sample were mixed and sequenced by 250 bp paired-end sequencing on Illumina MiSeq.