GSM8864726: No treatment, unsorted population, rep 3; Homo sapiens; OTHER
Genomic DNA of the sorted and unsorted fixed cells were then extracted using the protocol described previously. The sgRNA library was amplified by a two-step PCR protocol for NGS. All gDNA of sorted samples were used to maximize the representations. For the unsorted/bulk samples, 22890 ng of gDNA, which is relevant to 1250 x representation, was used for PCR1. PCR1 and PCR2 products were purified using AMPure XP Bead-Based Reagent (Beckman Coulter, A63881) according to manufacturer's instructions. The CRISPR knockout pooled library was purchased from Addgene (Cat#1000000083) and amplified using the protocol provided by Addgene.