GSM6739112: plasmid control part 2; Homo sapiens; OTHER
At screen end point cells were harvested and gDNA was extracted with QIAamp DNA Blood Maxi Kit (Qiagen #51194). Genome weight was estimated based on measured ploidy of cells. The sequencing library was prepared using NEBnext® Ultra II Q5 Master Mix (NEB #M0544) and custom primers (forward i5: 5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT[N]cccttggagaaccaccttgTTG-3' with N=(0-8) nt stagger and reverse primer for multiplexing: 5'-CAAGCAGAAGACGGCATACGAGAT[8N]GTGACTGGAGTTCAGACGTG-3')  to have a balanced read sample. A barcode in the reverse primer was used for identification of the sequencing libraries. Libraries were gel purified and cleaned up. Libraries were balanced and quality was assessed with Bioanalyzer High Sensitivity DNA Kit, Agilent #5067-4626). R1 was used for sgRNA identification, R2 was used to read out barcode associated with each single sgRNA in the pool