GSM6364360: G166, day 0, rep 2; Homo sapiens; OTHER
Genomic DNA was extracted using DNeasy Blood & Tissue kit (Qiagen). A two-step PCR procedure was employed using KAPA HiFi HotStart polymerase (KAPA) to amplify sgRNA sequences from genomic DNA and to incorporate add Illumina adapters and multiplexing barcodes to sgRNA amplicons. Purified PCR products were quantified by Qubit dsDNA HS Assay Kit (Thermo Fisher) and KAPA Library Quantification Kit (Roche), pooled in equimolar quantities and sequenced on a NextSeq 500 instrument using a single-end 75bp kit (Illumina). OTHER: CRISPR screen