The essential and non-essential tiling screen with Hsu tracrRNA(2013) in A375 cells, dropout screen, replicate B
A375_311Cas9 cells were transduced in three biological replicates with the lentiviral library containing Hsu tracrRNA.  Puromycin was added 2 days post-transduction and maintained for 5 days to ensure complete selection for transduced cells. Upon puromycin removal, cells were passaged every 2-3 days for an additional 2 weeks at a minimum of 500x representation, at which point, 21 days post-transduction, cells were collected; genomic DNA from these cells was PCR amplified and sequenced.