Hub screening in K562 replicate 2 Day 30
1. infected with pgRNA library lentiviruses at an MOI of <0.3 (1000-1500x coverage of the library).2. 72 hours after infection, EGFP+ cells were selected by FACS (Day-0).3. further maintained at a minimum coverage of 1500x for 30 days.4. genomic DNA extraction, PCR amplification of sgRNA-coding sequence and high-throughput sequencing.