GSM8256302: A375, DMSO, TN control, replicate 4; Homo sapiens; OTHER
After a 72h recovery and expansion, the genomic DNA was extracted from cells (40x coverage over the GeCKO library) using the blood and cell culture midi kit (Qiagen, 13343). Amplification first step PCR was performed to sgRNA sequences using the Herculase II Fusion DNA polymerase (Agilent Technologies, 600675). A second PCR was performed to attach Illumina adaptors and barcode samples. Amplicons from the second PCR were extracted (Qiagen), analyzed using a Bioanalyzer High Sensitivity DNA Analysis Kit (Agilent), and quantified using a NEBNext Library Quant Kit for Illumina.