GSM7595816: IGROV-1 cellsCRISPR Brunello libray, uninfected; Homo sapiens; OTHER
Total genomic DNA from both virus-resistant and uninfected control cells was respectively extracted using QIAamp DNA Mini Kit. The inserted guide RNA sequences were retrieved from the genomic DNA by PCR amplification using the forward 5'-AATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCG-3' and reverse 5'-CTTTAGTTTGTATGTCTGTTGCTATTATGTCTACTATTCTTTCC-3' primers. The PCR products were further barcoded by an additional round of PCR amplification using primers containing illumina i5 and i7 barcodes. The PCR products were then gel purified and subjected to next-generation sequencing on illumina MiSeq platform using a custom sequencing primer.