Workflow schematic for an sgRNA library screen using 6-thioguanine (6-TG) selection
Workflow schematic for an sgRNA library screen using 6-thioguanine (6-TG) selection. After transduction of Cas9+ A375 cells with the sgRNA library, hygromycin selection, and expansion, the Cas9+/ sgRNA+ cells were split into two populations. A part of the Cas9+/sgRNA+ cell population was exposed to 6-TG, while the control population was expanded under normal culture conditions, without adding 6-TG. After expansion of both cell populations, genomic DNA was harvested and prepared for sequencing with the Guide-it CRISPR Genome-Wide sgRNA Library NGS Analysis Kit.