GSM5380363: TargetSeq_Sample_49; Homo sapiens; OTHER
72 hours after transfection, the genomic DNA was extracted from the transfected cells with QuickExtractTM DNA Extraction Solution (QE09050, Epicentre) Target genomic sequences were PCR amplified by high-fidelity DNA polymerase PrimeSTAR HS (Clonetech) with primer sets flanking examined gRNA target sites. The gRNA target sequences and PCR primer sequences are listed in Supplementary Table 2. Indexed DNA libraries were prepared by using the NEBNext Ultra II FS DNA Library Prep Kit for Illumina. After quantitated with Qubit High-Sensitivity DNA kit (Invitrogen), PCR products with different tags were pooled together for deep sequencing by using the Illumina Hiseq X Ten (2×150) at Omics Core of Bio-Med Big Data Center, Shanghai Institute of Nutrition and Health, Shanghai, China.