gDNA PCR amplicon of mus musculus: mdx4cv heart
The  off-target loci were first amplified by genomic DNA PCR using gene-specific primers with Illumina adapters. The first PCR products were purified using a commercial purification kit (Promega, Madison, WI, USA), diluted, pooled, and subjected to a second round PCR with primers including the index sequences. The final PCR products were electrophoresed on an agarose gel, showing a single sharp peak. The purified amplicons were pooled and sent for sequencing using a MiSeq nano-scale flow cell.