GSM7426853: A549sgNegTSGT1_1; Homo sapiens; OTHER
DNA was extracted using Qiagen's Blood Maxi Kit, sgRNA cassettes were PCR-amplified from genomic DNA (T0 and end points) using NEB Q5 DNA Polymerase in order to add deep sequencing adapters and sample barcodes. Finally, sgRNA compositions was analyzed by deep sequencing using HiSeq4000 SE65 technology (Illumina). The sgRNA library was synthetized as a single oligo pool (Twist Bioscience), PCR-amplified and cloned into the lentiviral vector pMB160, derived from pLG20 (gift from Luke Gilbert, UCSF).