NODEGenomicsWeiwei ZhangAMPLICONhttps://www.biosino.org/node/experiment/detail/OEX00012567HiSeq X TenPairedPCRY5['cell Line']['Homo sapiens']Generally, biotinylated primer was designed within 150 bp around Cas9-targeting site to achieve primer extension. Site-specific nested primer was designed for following amplification. All the PEM-seq libraries were sequenced by Illumina Hiseq. For off-target analysis, junctions proximal to break-site (±20 kb) were excluded and MACS2 callpeak was used to identify translocation enrich region. Off-target hotspots were defined to have less than 8 mismatches with on-target site and have more than 3 junctions at the presumable cutting site.NODEfalsePEM-seqPEM-seq construction and analysis for off-target, translocation and large deletion.2021-03-122021-03-11OEX00012567OEP00001824