NGS of HCT116 cells expressing dCas9-ZIM3(KRAB)-MeCP2(t): Day 21, Rep2
gDNA was collected from ~50 million cells per replicate using the Zhang Lab salt-based extraction method. Libraries were generated by PCR-amplification of 100ug of gDNA using ~50 parallel reactions, using primers annealing to lentiviral transgene sequences. Following pooling and purification, a second parallel PCR step, using the cleaned first amplification as a template, was used to append Illumina adapter sequences and barcodes for multiplexing.