Sequencing of genomic loci targeted by Cas9

NGS analysis was performed for gene editing experiments at the BFP reporter and at the TP53, CALD1, FANCM and SPRTN loci. Five days after transfection with Cas9, sgRNAs and/or DNA donors, and either empty control vector or vector expressing an engineered RAD18 variant (e18) cells were collected and their genomic DNA was isolated. Sequencing libraries were prepared and the samples were analyzed by next generation sequencing.