synthetic lethality screen, gene-trap mutagenesis in haploid cells, second replicate
Construction Protocol: HAP1 cells were mutagenized as described previously (Blomen et al., 2015). In summary, 6x10E7 cells were transduced with pGT gene-trap retrovirus. Mutagenized cells were passaged for 12 days prior to fixation in Fixation Buffer 1 (Becton Dickinson). Cells were stained with DAPI (1 ug/ml), and G1 haploid cells were sorted. Genomic DNA was isolated from 4x10E7 cells, and subjected to a linear amplification (LAM)-PCR protocol, and sequenced using the Genome Analyzer platform (Illumina).