Whole Genome CRISPR of UM cells: FAKi treatment
Genomic DNA was isolated using NucleoBond CB 500 (Takara Bio cat#740509) according to the manufacturers protocol. DNA concentration was measured using a Qubit assay. Amplification of sgRNAs for NGS was performed according to the Broad Institute's recommended protocol. The entirety of isolated genomic DNA was used for subsequent PCR, to ensure capturing the full representation of the libraries. PCR products were sequenced on a HiSeq4000 instrument (Illumina) (350M Reads).NGS read counts were processed, aligned, using PinAPL-Py (v2.9.2). Read counts were analyzed using Mageck-MLE (0.5.9.5) to identify enrichment or depletion of sgRNAs in treatment vs control samples