[1] Similar to LAM-PCR, it employs multiple PCRs to amplify regions of interest that contain a specific insert that preferentially integrates into double-stranded breaks.
As gene therapy is an emerging field, GUIDE-Seq has gained traction as a cheap method to detect the off-target effects of potential therapeutics without needing whole genome sequencing.
[1] This makes it critical to have a dsODN only condition that controls for errant and naturally occurring DSBs, and is required to use the GUIDE-seq bioinformatic pipeline.
The final product is a panoply of amplicons, describing the DSB distribution, containing indices for sample differentiation, p5 and p7 Illumina flow-cell adapters, and the sequences flanking the dsODN cassette.
[3] GUIDE-Seq has been shown to miss some off-targets, when compared to the genome-wide sequencing DIGENOME-Seq method, due to the nature of its targeting.