Genome-wide, unbiased identification of DNA double-stranded breaks (DSBs) enabled by sequencing (GUIDE-seq), relies on capture of double-stranded oligodeoxynucleotides into DSBs. The majority of identified sites were not detected by existing computational methods or chromatin immunoprecipitation sequencing (ChIP-seq). GUIDE-seq also identified CRISPR RNA-guided nucleases-independent genomic breakpoint ‘hotspots’. Also, GUIDE-seq revealed that truncated guide RNAs exhibit substantially reduced RNA-guided nucleases-induced, off-target DSBs (PMID: 25513782).