OnTarget_Cas9Minus
Equal amounts of the gDNA was used to amplify genomic regions flanking the on-target off-target nuclease binding sites for library construction. Next, PCR amplicons were purified using Ampure beads (Beckman Coulter), then subject to second round PCR to attach Illumina P5 adapters and sample-specific barcodes. The purified PCR products were pooled at equal ratio for single-end sequencing using Illumina MiSeq at the Zhang laboratory (UCSD). Two sets of experiments were prepared separately for each target (Cas9Minus and Cas9Plus), which the Cas9Minus sample was the negative control used to calculate background indel rate and Cas9 wasn't added.