GSM8179509: HAP1 cells, SV1_CTRLB; Homo sapiens; OTHER
Genomic DNA was extracted from cell pellets with the ISOLATE II Genomic DNA Kit (Bioline). Samples of gDNA (25 μg for controls and 5 μg for selected samples) were added to NEBNext High-Fidelity 2X PCR Master Mix (New England Biolabs) along with the following primers to bind the sgRNA region: NGSCRISPRv2F1 5'-GGACAGCACAGATCCAGTTTGGT- 3' and NGSCRISPRv2R1 5'-GAGCCAATTCCCACTCCTTTCAA-3'. A second PCR reaction with a staggered primer mix of P5 and P7 indexing primers unique to each sample was prepared. Reactions were isolated by gel electrophoresis. Samples were sequenced with Novogene