Cells transduced with control, single cell cloned into well B08 of Plate3, and PCR amplified with MCL1_F3 and MCL1_R3
MELJUSO cells were transduced with control cloned into a vector containing BE3.9max and puromycin resistance. Cells were selected with puromycin for 7 days. Single cell clones were isolated by FACS using a MoFlo Astrios Cell Sorter to generate four 96-well plates per guide (20 plates total); this sample corresponds to Plate3, Well B08. Clones were expanded for 7 days; this plate was then cultured in the absence of drug for 16 days. After this, cells were collected and lysed, and the target locus was PCR amplified using the F / R primers and sequenced.