GSM9461550: Doxy+_1; Homo sapiens; OTHER
Cells were harvested on day 18, and genomic DNA was extracted using the Quick-DNA Midiprep Plus Kit according to the manufacturer's instructions. All screening experiments were performed in duplicate using independent biological replicates. The sgRNA cassettes were PCR-amplified from genomic DNA using KOD Plus Neo polymerase  with primers containing Illumina-compatible adapters and unique barcodes. PCR products were purified via agarose gel electrophoresis and a gel extraction kit. Purified libraries were quantified by qPCR using the KAPA Library Quantification Kit, normalized to 1 nM, and sequenced on an Illumina NextSeq 500.