GSM4120080: S07_t4_rep2; Homo sapiens; OTHER
Genomic DNA was isolated from ~50 milliion cells per sub-library, per transduction replicate at each timepoint.  Frozen cell pellets were thawed and genomic DNA was isolated using the standard protocol (Qiagen Blood and Cell Culture Maxi Kit, c/n 13362). Each sgRNA subpool library readout was performed using two steps of PCR as described in Chen et al. Cell (2015).  Second PCR round products were purified using column-based cleanup (NEB Monarch PCR and DNA Cleanup Kit, c/n T1030S).  Libraries of each time point for each sub-library were internally controlled by uniquely barcoding and combining for sequencing on the same channel(s) of a sequencing run.  Diluted libraries were spiked in with whole-exome libraries and sequenced on HiSeq4000 instruments (Illumina).
GEO Accession
GSM4120080